Actions of a carbocyanine dye on calcium-dependent potassium transport in human red cell ghosts. 1. 3,3'-Diethylthiadicarbocyanine iodide (diS-C2-(5)), a fluorescent dye widely used to estimate membrane potentials, inhibits the Ca-dependent K transport system of human red cells and ghosts. It does not affect non-specific cation leaks, net chloride movements or the Ca-activated ATPase. 2. The nature of the inhibitory effect depends upon the conditions under which the dye is applied. When Ca-containing ghosts with 100 mM internal K are suspended in a choline medium at 37 degrees C, 0.3 microM-diS-C2-(5) causes virtually complete inhibition of K transport within 1 min (haematocrit 0.3%). Inhibition cannot be reversed by washing the ghosts to remove bound dye, although partial recovery occurs on subsequent incubation at 37 degrees C. 3. Inhibition is less marked when ghosts are suspended in solutions containing 100 mM-K, and is largely reversed by washing. The degree of inhibition varies with the dye concentration: 50% inhibition occurs with a free diS-C2-(5) concentration of 0.7 microM, and about 5 x 10(6) dye molecules are bound per ghost. 4. Changes in internal and external K concentrations, keeping internal Ca constant, have no effect on the dye concentration required for 50% inhibition of K efflux. 5. When internal Ca is varied, keeping internal and external K constant, the dye concentration needed for 50% inhibition of K efflux varies inversely with the K efflux in the absence of dye. That is, diS-C2-(5) is most effective as an inhibitor when K efflux is at a maximum. This suggests that the dye reacts preferentially with the transport system in the Ca-activated state. 6. The effects of several related molecules were examined, using Rb influx as a measure of K permeability. Inhibition is obtained in some cases, but is not directly related to the ability of the ghosts to bind the dyes. An anionic dye, diS-C3SO3--(5) inhibits Rb influx only when present inside the ghosts. 7. The implications of these findings are discussed.


The control of blood flow through human forearm muscles following brief isometric contractions. 1. The blood flow through the forearm was measured 2 sec after single, brief isometric hand-grip contractions. The tension and duration of those contractions varied from 10 to 100% of the maximal voluntary contraction (m.v.c.) and from 2 to 12 sec, respectively. 2. The blood flow increased linearly with tension up to about 60% m.v.c. but further increases in tension, up to 100% m.v.c., did not elicit higher blood flows than were found at 60% m.v.c. The same relationship between tension and the resultant blood flow held for all durations of contractions, from 2 to 12 sec. The blood flow immediately after (2 sec) contractions at a given tension increased linearly with the duration of the contraction, from 2 to 12 sec. Maximal exercise blood flow was approached only in response to the longest contractions (12 sec) at tensions of 60% m.v.c. or higher. 3. Brief alterations (2--5 sec) of transmural pressure across blood vessels did not result in a significant change of blood flow, either in the resting forearm or when the vessels were dilated by brief, isometric contractions. When the tension was applied or released either rapidly or gradually ('ramp' contractions) there was no correlation between the rate of change of stretch on arterial vessels and the resultant blood flow. However, there was a direct relationship between a force--time integral (duration of contraction x peak tension) and blood flow. All these results make it clear that changes in blood flow in the forearm elicited by brief isometric contractions are not the result of a myogenic reflex but are metabolically induced. 4. Successive contractions exerted at 60% m.v.c. for 4 sec induced a blood flow of 21.2 +/- 1.6 ml.min-1.100 ml.-1 when a rest interval of 8 sec was allowed between the contractions. Blood flows remained constant at this submaximal level, even when muscular fatigue was induced, and when there was an accompanying large increase in blood pressure. 5. Isometric muscular activity by the contralateral arm which resulted in fatigue, associated with a large increase in mean blood pressure, did not alter the level of vasodilation that was induced by brief, isometric contractions in the 'test' arm. 6. It is suggested that the vasodilatation in response to intermittent isometric contractions is the result of metabolic vasodilatation of distal segments and continued sympathetic vasoconstriction of the proximal segments of the forearm vascular bed.


The change in the load-sustaining ability and in the series elasticity in Mytilus smooth muscle during isotonic shortening. 1. The change in the ability to sustain a load and the change in the series elasticity in the anterior byssal retractor muscle (a.b.r.m.) of Mytilus edulis during isotonic shortening was studied by recording the length changes following step changes in load. 2. When a load of 0.7--0.9 P0 WAs applied after a period of isotonic shortening under a small load (0.05--0.1 P0), the muscle fibres showed continuous isotonic lengthening, indicating a reduction in the ability to sustain a load during isotonic shortening. 3. Following the application of a load of 0.3--0.6 P0 during isotonic shortening under a small load, the fibres exhibited a transient isotonic lengthening before starting to shorten isotonically, indicating some degree of restoration in the load-sustaining ability after the step increase in load. 4. No appreciable reduction in the load-sustaining ability was observed during isotonic shortening under a large load (more than 0.7 P0). 5. The load--extension curves of the series elasticity determined during isotonic shortening were found to be scaled down roughly in proportion to the isotonic load. 6. The stiffness of the muscle fibres during the isotonic shortening approached a certain finite value, when the isotonic load tended to zero. If the stiffness was measured during the development of isometric tension, the stiffness--isometric tension curve extrapolated towards the origin. 7. High-speed cinematography during the step change in load indicated a fairly uniform distribution of the series elasticity along the length of the preparation. 8. These results are discussed in relation to the sliding filament model of muscle contraction.


The effects of tenotomy upon the contraction characteristics of motor units in rabbit soleus muscle. 1. Isometric contractions of tenotomized rabbit solei have been compared with a group of unoperated control muscles. A large decrease in the tension developed by the tenotomized muscle was accompanied by a slight shortening of the twitch contraction time, and a larger, progressive, reduction in the time to half relaxation.2. A comparison of motor units obtained from muscles which had been tenotomized for six weeks with those of the control muscles showed a large reduction in the range of contraction and half relaxation times of the units from the tenotomized muscles.3. The mean motor unit tension (expressed as a percentage of the whole muscle tension) was similar for both those units from the control muscles and those from muscles which had been tenotomized for six weeks, indicating a uniform atrophy of motor units within the tenotomized muscles.4. It was concluded that the change in the pattern of motor unit contraction times was not the result of a process of differential atrophy favouring the preservation of the faster contracting motor units.5. A correlation between axon conduction velocity and both the speed of contraction and the size (tension) of the motor units was demonstrated in the control muscles. Following tenotomy the relationship between axon conduction velocity and motor unit tension was lost.


Descending influences on the responses of spinocervical tract neurones to chemical stimulation of fine muscle afferents. 1. In cats, extracellular micro-electrode recordings were made from axons of the spinocervical tract (s.c.t.) in both the decerebrate state and during cold block of the spinal cord (reversible spinal state) to examine the effects of intra-arterial injection of algesic agents (bradykinin, potassium, 5-hydroxytryptamine) into the gastrocnemius-soleus (g.s.) muscle on the discharge behaviour of s.c.t. neurones.2. In the decerebrate state without cooling the spinal cord 13% of the cells (eleven out of eighty-three) responded to intra-arterial injection of bradykinin, 33% (twenty-two out of sixty-nine) to 5-hydroxytryptamine, and 38% (thirty-five out of ninety-one) to potassium injection.3. The general time course and the latency of the responses of s.c.t. cells induced by injection of pain-producing substances into the g.s. muscle reflect in many respects the activations of g.s. group III and group IV primary afferent units studied previously.4. For twenty-seven s.c.t. neurones the period of recording was long enough to record the responses of the same cell to injections of algesic agents in both the decerebrate and the reversible spinal state. In the reversible spinal state 83% (nineteen out of twenty-three) of the s.c.t. neurones tested with all the three substances responded to at least one of the algesic agents. In the decerebrate state the percentage was lower (39%).5. Reversible spinalization led not only to a significant increase in the number of s.c.t. neurones responding to the algesic agents used but also to an increase in the magnitude of the chemically induced responses.6. The mean latency of the responses of neurones that were activated in both preparations were shorter in the reversible spinal state than in the decerebrate state.7. Control experiments showed that the responses to bradykinin and potassium were entirely due to the nervous outflow from the g.s. muscle. In contrast, intra-arterially applied 5-hydroxytryptamine influenced the s.c.t. cells via unknown additional sites of action.8. The results indicate that muscular group III and/or group IV units excitable by algesic substances do project on to neurones of the spinocervical tract. Furthermore it is concluded that the responses of s.c.t. neurones to activation of fine muscle afferents by algesic agents are subject to a descending control similar to the well known descending modulation of their responsiveness to cutaneous input. Therefore, in addition to serving as a cutaneous pathway the spinocervical tract may take part in muscular nociception.


Nonlinear spatial summation and the contrast gain control of cat retinal ganglion cells. 1. We studied how responses to visual stimuli at spatially separated locations were combined by cat retinal ganglion cells. 2. The temporal signal which modulated the stimuli was a sum of sinusoids. Fourier analysis of the ganglion cell impulse train yielded first order responses at the modulation frequencies, and second order responses at sums and differences of the input frequencies. 3. Spatial stimuli were spots in the centre and periphery of the cell's receptive field. Four conditions of stimulation were used: centre alone, periphery alone, centre and periphery in phase, centre and periphery out of phase. 4. The effective first order response of the centre was defined as the response due to centre stimulation in the presence of periphery stimulation, but independent of the relative phases of the two regions. Likewise, the effective first order response of the periphery was defined as the response due to periphery in the presence of centre stimulation, but independent of the relative phases of the two regions. These effective responses may be calculated by addition and subtraction of the measured responses to the combined stimuli. 5. There was a consistent difference between the first order frequency kernal of the effective centre and the first order kernel of the centre alone. The amplitudes of the effective centre responses were diminished at low frequencies of modulation compared to the isolated centre responses. Also, the phase of the effective centre's response to high frequencies was advanced. Such non-linear interaction occurred in all ganglion cells, X or Y, but the effects were larger in Y cells. 6. In addition to spatially uniform stimuli in the periphery, spatial grating patterns were also used. These peripheral gratings affected the first order kernal of the centre even though the peripheral gratings produced no first order responses by themselves. 7. The temporal properties of the non-linear interaction of centre and periphery were probed by modulation in the periphery with single sinusoids. The most effective temporal frequencies for producing non-linear summation were: (a) 4-15 Hz when all the visual stimuli were spatially uniform, (b) 2-8 Hz when spatial grating patterns were used in the periphery. 8. The characteristics of non-linear spatial summation observed in these experiments are explained by the properties of the contrast gain control mechanism which we have previously postulated.


Segmental and supraspinal input to cells of origin of non-primary fibres in the feline dorsal columns. 1. The synaptic input to ascending tract cells with axons in the dorsal columns was investigated using intracellular recording. 2. E.p.s.p.s evoked by stimulation of the lateral funiculus were analysed to test for the possibility of collateral connexions between spino-cervical tract cells and dorsal column cells. Three groups of fibres were found to contribute to such e.p.s.p.s: fibres which terminated or originated between spinal segments C3-4 and C1, or Th9 and C3-4 and cortico-spinal tract fibres. The latencies and thresholds of e.p.s.p.s evoked by stimulation of the first group of fibres were compatible with their origin via axon collaterals of spino-cervical tract cells. The occurrence of these e.p.s.p.s in dorsal column cells which were disynaptically excited from cutaneous afferents further corroborated this possibility. 3. E.P.S.P.S of specifically cervical origin were also found in some other neurones in the dorsal horn, probably segmental interneurones, but were absent in spinocervical tract cells. 4. Convergence of group I muscle afferents (possibly both group Ia and group Ib) and cutaneous afferents was found in about 50% of the dorsal column cells. The shortest latency e.p.s.p.s from cutaneous and group I afferents were evoked with segmental delays indicating monosynaptic and disynaptic coupling. 5. I.p.s.p.s were evoked from cutaneous and group I muscle afferents in either the same or different nerves as those from which the e.p.s.p.s were elicited. Excitatory potentials were, however, dominating.


Three-micro-electrode voltage clamp experiments in calf cardiac Purkinje fibres: is slow inward current adequately measured? 1. The three-micro-electrode voltage clamp method (Adrian, Chandler & Hodgkin, 1970) was adapted for the study of regenerative inward currents in cardiac muscle. The adequacy of measurements of slow inward current in cardiac Purkinje fibres was assessed. 2. Membrane current density is reported simultaneously by total applied current (IT), along with a longitudinal voltage difference signal (delta V), recorded between two intracellular micro-electrodes. 3. Non-linear cable calculations show that delta V is a more faithful measure of membrane current density than IT as peak inward current increases. Quantitative agreement between delta V and IT only occurs when both signals report the membrane characteristics that would be obtained with an ideal longitudinal space clamp. 4. Agreement between delta V and IT is thus a useful criterion for satisfactory experimental measurements which we applied to the slow inward current. This component was elicited by depolarizing steps from a holding potential near =45 mV in the presence of tetrodotoxin. The IT signal was compared directly with delta V/R, where R is an effective longitudinal resistance that was experimentally determined. 5. delta V/R and IT showed very good agreement in both peak amplitude and time course at all potentials studied. 6. Radial non-uniformity during the measured peak slow inward current was estimated by calculations assuming clefts 200 A wide with a uniform distribution of ionic channels. The calculated voltage span from surface to centre was always less than 5 mV, and the measured I-V characteristics showed little distortion. 7. In another check, I-V characteristics and slow response membrane action potentials were compared. The measured peak current showed good agreement with the product (total preparation capacitance) x (rate of rise). 8. The experimental and theoretical analysis suggest that the measurements of slow inward current are a good approximation to genuine membrane properties.


Tetanus toxin and synaptic inhibition in the substantia nigra and striatum of the rat. 1. The effects of tetanus toxin were determined on GABA-mediated synaptic inhibition of substantia nigra neurones evoked by striatal stimulation and on the presumed dopamine- and 5-hydroxytryptamine-mediated synaptic inhibition of striatal neurones evoked by nigral and dorsal raphe nucleus stimulation, respectively, in the urethane-anaesthetized rat. 2. Following an intranigral injection of tetanus toxin, striatal-evoked inhibition of substantia nigra neurones, which is sensitive to bicuculline, was rapidly abolished. This effect was not accompanied by any significant change in the responses of nigral neurones to ionophoretically administered GABA or other putative neurotransmitters and thus indicates a presynaptic site of action of the toxin. 3. The rate of onset of action of the toxin in the substantia nigra was extremely rapid (1-4 min) and appeared to be related to the rate of activation of the inhibitory pathway. 4. Injections into the substantia nigra of tetanus toxin neutralized with antitoxin had no significant effect on striatal-evoked inhibition in the substantia nigra. 5. Injections of tetanus toxin into the striatum failed to influence the inhibition of striatal neurones evoked by stimulation of the ipsilateral substantia nigra or the dorsal raphe nucleus, suggesting that tetanus toxin does not impair monoamine-mediated inhibition in the central nervous system. 6. Synaptic excitation which preceded substantia-nigra-evoked inhibition in striatal neurones and which occasionally preceded striatal-evoked inhibition in nigral neurones was also unaffected by tetanus toxin. 7. It is suggested that tetanus toxin selectively abolishes GABA-mediated synaptic inhibition in the central nervous system and may be a useful tool in the identification of such synaptic inhibitory mechanisms.


Electrical properties associated with wide intercellular clefts in rabbit Purkinje fibres. 1. Rabbit Purkinje fibres were studied using micro-electrode recordings of electrical activity or a two-micro-electrode voltage clamp. Previous morphological work had suggested that these preparations offer structural advantages for the analysis of ionic permeability mechanisms. 2. Viable preparations could be obtained consistently by exposure to a K glutamate Tyrode solution during excision and recovery. In NaCl Tyrode solution, the action potential showed a large overshoot and fully developed plateau, but no pacemaker depolarization at negative potentials. 3. The passive electrical properties were consistent with morphological evidence for the accessibility of cleft membranes within the cell bundle. Electrotonic responses to intracellular current steps showed the behaviour expected for a simple leaky capacitative cable. Capacitative current transients under voltage clamp were changed very little by an eightfold reduction in the external solution conductivity. 4. Slow current changes attributable to K depletion were small compared to those found in other cardiac preparations. The amount of depletion was close to that predicted by a cleft model which assumed free K diffusion in 1 micron clefts. 5. Step depolarizations over the plateau range of potentials evoked a slow inward current which was resistant to tetrodotoxin but blocked by D600. 6. Strong depolarizations to potentials near 0 mV elicited a transient outward current and a slowly activating late outward current. Both components resembled currents found in sheep or calf Purkinje fibres. 7. These experiments support previous interpretations of slow plateau currents in terms of genuine permeability changes. The rabbit Purkinje fibre may allow various ionic channels to be studied with relatively little interference from radial non-uniformities in membrane potential or ion concentration.


The initiation and maintenance of bradycardia in a diving mammal, the muskrat, Ondatra zibethica. 1. The cardiac and respiratory responses shown by muskrats in both unrestrained and restrained dives have been compared with responses elicited by stimulation of a number of cardio-depressant receptor inputs, in an attempt to determine which are most important in initiating and maintaining diving bradycardia. 2. In unrestrained voluntary dives heart rate fell from 310 +/- 3 to 54 +/- 3 beats min-1 in 1 to 2 sec, which was significantly below that seen in dives by restrained unanaesthetized or anaesthetized animals. 3. Pouring water on the external nares during maintained artificial ventilation caused heart rate to decline to 76 +/- 12 beats min-1 after 1 sec. Flowing water through the internal nares caused apnoea, in the expiratory position, and bradycardia within one third of a second. Heart rate fell to 20 +/- 2 beats min-1, 1 sec after the start of water flow. Substituting saline for water reduced both the apnoeic and cardiac responses. Bilateral section of the maxillary branch of V and the inferior laryngeal (X) nerves completely abolished the cardiac and respiratory response to water flow. 4. Artificial ventilation throughout periods of nasal stimulation with water or saline reduced the bradycardia, although even the saline driven response could not be completely abolished. Lung deafferentation eliminated any direct effect of artificial ventilation on heart rate during nasal stimulation. 5. Lung deflation caused bradycardia within 0.97 +/- 0.17 sec, heart rate falling from 268 +/- 7 to 59 +/- 4 beats min-1. Bradycardia also occurred during maintained lung inflation but it was delayed for a period which varied from 6.8 +/- 1.8 sec at an inflation pressure of 0.5 kPa to 35 +/- 7 sec at 1.5 kPa. 6. Bradycardia caused by nasal water flow or lung deflation was unaffected by bilateral section of the sinus nerve. 7. Artificial ventilation of paralysed muskrats with 5% CO2 in N2 caused bradycardia when Pa, O2 reached 8.4 +/- 0.8 kPa and heart rate declined to 76 +/- 7 beats min-1 at 4 kPa. Bilateral section of the sinus nerve delayed bradycardia until Pa, O2 reached 4.5 +/- 0.5 kPa. 8. These results suggest that the cardiac response to submergence could be the expression of input from three groups of receptors, nasal, lung and carotid chemoreceptors, although it is not clear how they interact with one another to generate the cardiac responses displayed by unrestrained animals during submergence.


Pancreatic acinar cells: effects of micro-ionophoretic polypeptide application on membrane potential and resistance. 1. Acinar cell membrane potential and resistance were measured from superfused segments of mouse pancreas, in vitro, using intracellular glass micro-electrodes. One or two extracellular micropipettes containing caerulein, bombesin nonapeptide (Bn) or acetylcholine (ACh) were placed near to the surface of the impaled acinus. The secretagogues were ejected rapidly from the micropipettes by ionophoresis.2. Each secretagogue evoked a similar electrical response from the impaled acinar cell: membrane depolarization and a simultaneous reduction in input resistance. The duration of cell activation from caerulein ionophoresis was longer than that observed for ACh and Bn. The cell response to the peptide hormone applications could be repeated in the presence of atropine.3. The minimum interval before the onset of cell depolarization after caerulein ionophoresis was determined. Values ranged between 500 and 1000 msec. The minimum latencies after Bn ionophoresis were 500-1400 msec.4. With two electrodes inserted into electrically coupled acinar cells, direct measurements of the caerulein and Bn null potentials were made. At high negative membrane potentials an enhanced depolarization was evoked by caerulein ionophoresis. At low negative membrane potentials the caerulein stimulation produced a diminished depolarization, and at membrane potentials less than - 10 mV acinar cell hyperpolarizations were observed. A similar series of responses was obtained in experiments where Bn ionophoresis was used. The caerulein and the Bn null potentials were always contained within - 10 to - 15 mV.5. The results describe the almost identical electrical response of acinar cells to stimulation by ACh, caerulein and bombesin. All three secretagogues have similar null potentials and latencies of activation on acinar cells. The bombesin latency responses appear as short as those measured for caerulein and provide electro-physiological evidence that Bn acts directly on acinar cells. The findings support the hypothesis that ACh, caerulein and Bn, though acting on different receptors, evoke the observed changes in electrical properties of acinar cell membranes, through a common pathway.


The effects of theophylline and choleragen on sodium and chloride ion movements within isolated rabbit ileum. 1. Theophylline (10 mM) and choleragen change the direction of net Cl- movements across rabbit ileum, in the short-circuit current condition, from absorption to secretion. The specific activity ratio R of Cl- tracers within the tissue coming from mucosal and serosal solutions respectively is increased, which is consistent with an increase in Cl- exchange flux across the mucosal border. 2. Net Na+ movement is also changed from net absorption to secretion by theophylline and choleragen; the specific activity ratio R of Na+ tracers is raised by theophylline. Because of the large paracellular component to transepithelial Na+ movements, an increase in Na+ exchange flux across the mucosal border is not detected. 3. 2,4,6-Triaminopyrimidine (20 mM) which has been previously shown to block paracellular Na+ movements, blocks both the theophylline and choleragen-dependent reversal of net Na+ movement by preventing the decrease in m-s Na flux. The theophylline-dependent increase in the ratio R of Na+ is still present, and is consistent with an increase in Na+ exchange flux across the mucosal border--unmasked by removal of the paracellular flux components. 4. Ouabain (0.1 mM) abolishes net absorption of Na+ and Cl- in control and net secretion of Na+ and Cl- in theophylline-treated tissue. Ouabain does not affect the theophylline-dependent increase in Cl- exchange across the mucosal border. 5. Replacement of Ringer Cl- with SO24- or Na+ by choline prevents the effects of theophylline and choleragen on Na+ and Cl- fluxes respectively. 6. Ethacrynate (0.1 mM) prevents the theophylline-dependent effects on net Na+ movement. Raising ethacrynate to 0.2 mM abolishes the effects of theophylline on Cl- exchange. An interpretation of these results is that theophylline and choleragen raise the Cl- permeability of the brush border. This increases NaCl leakage from the hypertonic lateral intercellular space into the mucosal solution thereby causing secretion. The selective action of triaminopyrimidine and ethacrynate (0.1 mM) on Na+ flux indicates that Na+ and Cl- move via separate transport pathways across the mucosal border.


Fluid movements across rabbit ileum coupled to passive paracellular ion movements. 1. Theophylline (10 mM) and choleragen (1 x 10(-6) g ml.-1) abolish net fluid absorption by everted sacs of rabbit ileum. Triaminopyrimidine (20 mM) and ethacrynate (0.1 mM) prevent this inhibition of net fluid movement. Replacing Ringer Cl- with isethionate prevents the theophylline-dependent decrease in fluid absorption also. 2. Ouabain (0.1 mM) abolishes net fluid movements in both control and theophylline-treated tissue. 3. With ouabain present, hypertonic NaCl (200 mM) in the mucosal solution causes net fluid secretion (serosal-mucosal flux). With theophylline added to both the mucosal and serosal solution, net fluid absorption (mucosal-serosal flux) is observed (P less than 0.001). Triaminopyrimidine (20 mM), or ethacrynate (0.1 mM), or replacement of Ringer Na+ with choline, or Ringer Cl- with isethionate all prevent the theophylline-induced reversal of osmotic flow. 4. Theophylline increases passive net flux of Na+ and Cl- from mucosal solution containing hypertonic (200 mM) NaCl+ ouabain (0.1 mM) across sheets of ileum into serosal solution containing mannitol Ringer + ouabain. The increased passive Na+ flux is blocked by triaminopyrimidine and the increased Na+ and Cl- fluxes are blocked by ethacrynate (0.1 mM). 5. The suggested route of increased NaCl leakage is via the paracellular pathway as it is inhibited by triaminopyrimidine. The increase, itself, is a consequence of the increased passive permeability of the mucosal border to Cl-, induced by theophylline or choleragen. Water is apparently electro-osmotically coupled to the paracellular Na+ leakage (100 mole water mole-1 Na+), hence increased passive leakage reverses osmotic flow. In active tissue the lateral intercellular space contains hypertonic NaCl, and hence increased leakage of NaCl across the tight-junction in theophylline or choleragen-treated tissue gives rise to net fluid secretion.


Electrophysiological and electron-microscopical correlations with fluid and electrolyte secretion in rabbit ileum. 1. The effects of theophylline and triaminopyrimidine on the passive permeability of Na+ and Cl- across sheets of rabbit ileum treated with 0.1 mM-ouabain were examined by determining the NaCl: mannitol dilution potentials, K+:Na+; choline: Na+ and SO24-:Cl- biionic potentials. The results indicate (a) that triaminopyrimidine reduces paracellular Na+ and K+ permeability without affecting Cl- permeability and (b) that theophylline increases Cl- permeability without affecting Na+ permeability and (c) that neither theophylline nor triaminopyrimidine interfere with each other's action. This is further evidence consistent with separate routes for paracellular Na+ and Cl- movement. 2. Electrical resistance changes across sheets of actively transporting rabbit ileum were measured as a function of time in various conditions. Theophylline has a biphasic effect on resistance. Initially it decreases resistance from 56 omega cm2 (control) to 40 omega cm2. In the ensuing 30 min, resistance rises to 50 omega cm2; whereas it falls to 30 omega cm2 in controls. When theophylline is present, with triaminopyrimidine, or with galactose, no secondary rise in tissue resistance occurs. The initial decrease in resistance is consistent with a theophylline-dependent increase in Cl- conductance and the secondary rise in resistance may be attributed to collapse of the lateral intercellular space following leakage of NaCl and fluid into the mucosal solution. 3. Electron microscopy of glutaraldehyde-fixed tissue confirms the above views, since, with theophylline present, the lateral intercellular spaces are collapsed and with both triaminopyrimidine and theophylline, or both theophylline and 20 mM-galactose present the spaces remain open. 4. It is shown in the Discussion that the theophylline- or choleragen induced increase in passive Cl- permeability of the mucosal border is the only requirement necessary to explain the increase in electrogenic Cl- secretion, the increase in short-circuit current, as well as neutral secretion of NaCl and net fluid secretion.


The response characteristics of an inhibitory binocular interaction in human vision. 1. The contrast threshold level for visual detection of a linear grating consisting of parallel light and dark bars is increased by adaptation to a high contrast grating of spatial characteristics similar to those of the test grating (e.g. Blakemore & Campbell, 1969). This so-called contrast threshold elevation effect is significantly reduced if two adaptation gratings, with appropriate different spatial characteristics, are presented one to either eye (Ruddock & Wigley, 1976). We have studied the contrast threshold elevation effect obtained with a test and an adaptation grating presented to one eye and a second adaptation grating, referred to as the conditioning grating, presented to the other. 2. Preliminary data are presented for three subjects with normal stereoscopic vision. In each case, the contrast threshold elevation effect for a pair of spatially identical test and adaptation gratings is significantly reduced by a conditioning grating of spatial frequency in the range 1-5 c/deg. 3. Reduction in the contrast elevation effect is observed whether the conditioning and adaptation gratings are of the same or of different wave-lengths and the effect of the conditioning grating increases to a steady-state level over a period of some 30 sec following onset of its presentation. 4. Measurements were made with a 5 deg diameter circular test grating presented at retinal locations up to 8 deg in the horizontal and vertical meridians from a foveal fixation point. It was found that in the horizontal meridian, the amplitude of the suppression effect associated with the conditioning grating falls as the displacement angle of the test field from the fixation point increases. For displacements in the vertical meridian, however, the amplitude of the suppression effect is virtually independent of the retinal location of the test field. 5. Under experimental conditions which yield suppression of the contrast threshold elevation effect for subjects with normal stereoscopic vision, no suppression was found for three subjects who possessed neither global nor fine stereoscopic vision. Results for a subject who possessed fine, but not global stereoscopy, did, however, show the suppression effect.


Electromechanical noise in atrial muscle cells of the carp: a possible ionic feed-back mechanism. 1. Spontaneous electrical fluctuations (+/- 0.2-0.4 mV) were recorded in 'quiescent' atrial fibres of the carp. 2. The noise decreased in acetylcholine, small hyperpolarization and EGTA. 3. The noise increased (to +/- 1-2 mV) and became more synchronous in K+-free Ringer and in ouabain. 4. Large voltage fluctuations (+/- 1-2 mV) were accompanied by a fine mechanical tremor, indicating intracellular [Ca2+] fluctuations. 5. Spectral analysis showed a clear resonant frequency at about 1 Hz, indicating that the noise cannot result from the random switching of independent ionic channels. 6. We propose that the intracellular [Ca2+] and the membrane K+-conductance are involved in a feed-back loop which can oscillate and produce the electromechanical noise. The frequency of oscillation is determined by the relatively slow diffusion of Ca2+ from the intracellular reservoir to the surface membrane.


Hormones, feeding and temperature in sheep following cerebroventricular injections of neurotransmitters and carbachol. 1. The neural mechanisms involved in the control of prolactin and growth hormone secretion, body temperature and feeding in castrated male sheep were investigated by the lateral cerebroventricular injection of several putative neurotransmitters and carbachol. 2. Feeding was inhibited by 10 and 20 microgram carbachol and by 400 microgram histamine. 100 microgram Noradrenaline stimulated, but 200 microgram inhibited, feeding; however, both  results were non-significant compared with controls. 3. 20 microgram Carbachol elicited a highly significant rise in plasma growth hormone, suggesting a cholinergic component in the neural control of growth hormone in sheep. Noradrenaline, dopamine and 5-hydroxytryptamine, but not melatonin, significantly depressed plasma prolactin concentration indicating a similar neural control for the hormone's release in sheep as in other species. 4. Deep body temperature appeared unaffected by any ofthe injected drugs. 5. The possible interrelationships between feeding behaviour and the circulating levels of growth hormone in ruminants is discussed. 6. A method for the radioimmunoassay of ovine growth hormone is described and validated.


Effect of acetylcholine stimulation on calcium and sodium uptake by the isolated rat pancreas. 1. The role of Ca2+ and Na+ ions in 'stimulus-secretion' coupling in the isolated uncinate pancreas of 4-week-old rats has been examined using radioisotope tracer techniques. The amount of 45Ca2+ and 22Na+ (mumole/g wet wt.) taken up by unstimulated glands was compared to that of glands in which amylase release was stimulated by ACh(10(-5) M) at various incubation times. 2. The amount of 45Ca taken up by the glands within 60 min of incubation was not found to be increased by the presence of ACh(10(-5) M). In fact, during short incubation periods the 45Ca uptake was significantly less in the stimulated glands than the unstimulated glands. 3. Presence of ACh(10(-5) M) did cause a significant initial increase in 22Na uptake lasting up to 20 min from onset of stimulation. 4. These results indicate that the rise in intracellular Ca2+ concentration which is suggested by an increase in 45Ca efflux during the action of pancreatic secretagogues, is not a consequence of increased 45Ca uptake by the pancreas; but they indicate that an initial action of ACh could be to elicit an increase in Na+ influx and that Na+ ions are likely to be involved in the action of ACh on pancreatic acinar cells.


Phasic firing enhances vasopressin release from the rat neurohypophysis. 1. Isolated rat neural lobes were incubated in vitro and electrically stimulated to release vasopressin. The released vasopressin was assayed using a radioimmunoassay and there was a reasonably good correlation (r = 0.81) between results obtained with this assay and those obtained by bioassay with the rat blood pressure method.2. Regular stimulation at frequencies of 5, 10 and 20 Hz released progressively more vasopressin and the release could be blocked by addition of tetrodotoxin to the incubation medium.3. Stimulation with pulse patterns derived from tape recordings of phasically firing units in the supraoptic nucleus of dehydrated rats released more vasopressin than the same number of pulses regularly spaced in time. In the range 2-8 pulses/sec vasopressin release was related to the pulse frequency within the bursts (r = 0.90) and the number of short (< 100 msec) interpulse intervals (r = 0.92). Vasopressin released per pulse increased over the frequency range 3-6 pulses/sec, but above 6 pulses/sec vasopressin release per pulse tended to diminish.4. We conclude that phasic firing of vasopressin neurosecretory cells may enhance vasopressin release in vivo and that an important factor in determining release is the number of short interspike intervals.


Electrophysiological identification of two types of fibres in rat extraocular muscles. 1. The synaptic potentials and electrical properties of rat inferior rectus muscles were examined in vitro. 2. In most fibres the spontaneous synaptic activity consisted of typical miniature end-plate potentials which had a normal distribution of amplitudes and rather uniform time courses. Suprathreshold and maximal nerve stimulation evoked unitary end-plate potentials (e.p.p.s). The synaptic activity of these fibres could be recorded only in the innervation zone of the muscle. These fibres were identified as being focally innervated. 3. Focally innervated fibres gave action potentials upon direct and indirect stimulation. They had an effective resistance (Reff)  of 1.62 +/- 0.22 M omega (mean +/- S.E., twenty-two fibres) and a time constant (tau m) of 3.8 +/- 0.4 msec (twenty-one fibres). Voltage-current curves in control saline were linear between membrane potentials of -50 to -140 mV. 4. In a small number of fibres the spontaneous synaptic activity consisted of miniature small-nerve junction potentials which had a skewed distribution of amplitudes with predominance of smaller voltages and time courses with a wide range of variation. Nerve stimulation evoked composite small-nerve junction potentials (s.j.p.s) which could be resolved into unitary components by varying the strength of stimulation. S.j.p.s had a higher threshold than e.p.p.s. Synaptic potentials could be recorded outside the innervation zone, at various sites along the muscle length. These fibres were recognized as being multiply innervated with polyneuronal innervation. 5. Multiply innervated fibres lacked action potentials had a large Reff of 6.0 +/- 1.1 M omega (six fibres) and a prolonged tau m of 29.8 +/- 4.8 msec. Reff show a moderate decrease to hyperpolarization and a rather large decrease to depolarization which denote, respectively, the presence of anomalous and delayed reactification. 6. It is concluded that rat extraocular muscles contain at least two populations of muscle fibres that in terms of synaptic activity and electrical properties are comparable to twitch fibres of other mammalian muscles and to slow or tonic fibres of amphibians.


Calcium-dependent potentials in the mammalian sympathetic neurone. 1. Intracellular recordings from post-ganglionic neurones of the rat superior cervical ganglion revealed two non-synaptic potentials dependent upon Ca2+, a hyperpolarizing afterpotential (h.a.p.) and a tetrodotoxin (TTX)-insensitive spike. 2. The h.a.p. followed regeneration discharge of the membrane potential in normal and TTX-containing Locke solution. 3. The h.a.p. appeared to arise from an increased K+ conductance because it was associated with a decrease in input resistance, reversed at -90 mV, and was proportional in magnitude to the extracellular K+ concentration. 4. Tetraethylammonium (TEA) and 4-aminopyridine (4-AP) apparently antagonized a voltage-sensitive K+ conductance because they broadened the action potential. However, these substances reduced only slightly the peak amplitude and earliest phases of the h.a.p. 5. The TTX-insensitive spike was most apparent when TEA was present and was invariably followed by an h.a.p. with a magnitude proportional to that of the spike. 6. The magnitude of the h.a.p. and the TTX-insensitive spike was directly proportional to the external Ca2+ concentration and was antagonized by Co2+ and Mn2+ in a dose-dependent fashion. 7. In normal Locke solution, Ba2+ antagonized the h.a.p. and allowed the neurone to sustain discharge during prolonged depolarization. In Locke solution containing TTX and TEA, Ba2+ reduced the magnitude of the h.a.p. but greatly increased the duration of the TTX-insensitive spike. 8. The h.a.p. was not significantly affected by altering external Cl- concentration and the TTX-insensitive spike was not reduced by altering external Na+ concentration. 9. It is concluded that the post-ganglionic neurone supports a regenerative Ca2+ conductance mechanism which in turn triggers an increased K+ conductance. The h.a.p. appears to result from outward K+ current in both a Ca2+ and voltage-dependent fashion.


Non-parallel enzyme secretion from rat pancreas in vitro studies. 1. The relative variations of rat pancreatic amylase versus lipase and chymotrypsinogen secretions have been studied in vitro with the help of two different techniques: in situ organ perfusion and incubation of pancreatic lobules. 2. In experiments on the perfused pancreas, with 8 C u.kg-1 hr-1 secretin added to the perfusion fluid, cholecystokinin-pancreozymin (8 ID u.kg-1) or pilocarpine (15 mg kg-1) both resulted in a significant change in the enzyme proportions in the juice. 3. In experiments on pancreatic lobules, the addition to the incubation medium of secretin (10(-7) M), alone or associated with cholecystokinin-pancreozymin (8 x 10(-7) M) or pilocarpine (10(-4) M) did not induce any change in the enzyme proportions in secretion. 4. It was concluded that the non-parallelism between enzyme secretions can occur in the rat upon pancreozyminic or cholinergic stimulation in vitro as well as in vivo (Dagorn, 1978) provided basal protein output is low enough. This is not the case when the tissular integrity of the pancreas is lost, such as in experiments on lobules. 5. This work confirms that pancreatic secretion derives from two intrapancreatic pools of different enzymatic composition, and gives a possible explanation for some discrepancies from the literature on the existence of non-parallel secretion.


Potassium activity in photoreceptors, glial cells and extracellular space in the drone retina: changes during photostimulation. 1. A double-barrelled potassium-sensitive micro-electrode was developed that was fine enough to record intracellular electrical potentials and potassium activities (aK) in the drone retina. 2. aK was measured in the photoreceptor cells, in the pigment (glial) cells, and in the extracellular space, in the superfused, cut, retina. The effect of photostimulation was studied: 20 msec light flashes, intense enough to evoke receptor potentials of maximum amplitude were presented, 1/sec, in a train lasting about 2 min. 3. In photoreceptors with membrane potentials greater than or equal to 50 mV aK in the dark was 79 mM, S.D. = 27 mM, n = 11. During photostimulation aK fell by 21.5 +/- 9.5 mM with a half-time of 30 +/- 22 sec. (A tentative conversion from activities to free concentrations can be made by taking the activity coefficient as 0.70 its value in the Ringer solution). 4. In pigment cells with membrane potentials greater than or equal to 50 mV, aK in the dark was 52  mM, S.D. = 13 mM, n = 11. During photostimulation aK increased by 14 +/- 5 mM. 5. In the extracellular space aK increased during photostimulation with a mean half-time of less than 1.3 sec to a maximum (mean value 14 mM, S.D. = 8.4 mM, n = 22), and then fell to a plateau. 6. It is estimated from the anatomy that the photoreceptors occupy approximately 38% of the total volume of the retina, the pigment cells 57%, and extracellular space 5%. Hence, it seems possible that during photostimulation nearly all the net loss of potassium from the photoreceptors is temporarily stored in the pigment cells. 7. Recordings were made in the extracellular space of the intact animal by passing the electrode through a hole in the cornea. The mean aK in the dark was 7.7 mM, S.E. = 0.4 mM, n = 22. In the superfused retina, aK in the dark was 6.3 mM, S.E. = 0.7 mM, n = 22, even though aK in the Ringer solution was 2.2 mM. Increasing the aK of the Ringer solution to 7.0 mM had no apparent effect on aK in the extracellular space at depths greater than 20 micron. 8. In the intact animal the amplitude and time course of the change in extracellular aK evoked by the standard pattern of photostimulation were within the range observed in the superfused preparation.


Desensitization of glutamate receptors on innervated and denervated locust muscle fibres. Depolarizations to L-glutamate, applied locally by microinophoresis to the extrajunctional membrane of locust extensor tibiae muscle fibres and measured either in current clamp or voltage clamp, increased in amplitude for equivalent doses of glutamate following chronic denervation of the muscle. 2. A two-pulse method was used to examine recovery from desentization of junctional and extrajunctional receptors. A 'response ratio', i.e. the amplitude of response to the second (test) of a pair of glutamate pulses over to the pulses. The 'response ratio' for extrajunctional depolarizations of innervated fibres increased exponentially with pulse interval, with a time constant of 15.6 +/- 4.7 sec (n = 11). Recovery of extrajunctional receptor populations from desensitization was accelerated after denervation. The recovery kinetics for responses from fibres 6-22 days after denervation were generally described by two exponential terms, with time constants in the range 0.5-10 sec which were inversely related to the glutamate sensitivity of the extrajunctional membrane. For junctional receptors on both innervated and denervated fibres the recovery kinetics were described by a single exponential with a time constant of 0.2-1 sec. 3. The results suggest that the increased extrajunctional glutamate sensitivity which occurs after denervation results from the 'appearance' of glutamate receptors with properties similar to those found at the post-junctional membrane on locust muscle fibres.


The importance of flow pulsatility for the rate of transvascular fluid filtration in lungs. 1. The rate of transvascular fluid filtration has been studied with a gravimetric technique in isolated perfused rabbit lungs during periods of elevated left atrial pressure (PLA). 2. Fluid filtration was expressed as the filtration coefficient, Kf (g/min x 100 g bloodless lung x mmHg PLA) and determined during alternately pulsatile and non-pulsatile perfusion in six zone III and three zone II/I lung preparations. Perfusion pattern was changed without interruption of flow. Mean in- and outflow pressures were kept constant. 3. In all the lungs it was found that Kf was higher during pulsatile than during non-pulsatile flow (P less than 0.01). Mean Kf (+/- S.E. of mean) for the zone III preparations was 0.42 (+/- 0.089) and 0.27 (+/- 0.057) for pulsatile and non-pulsatile perfusion, respectively. The corresponding figures for the zone II/I preparations were 0.11 (+/- 0.035) and 0.04 (+/- 0.030). 4. We suggest that the difference is due to a larger filtration area and/or a higher mean microvascular hydrostatic pressure during pulsatile than during non-pulsatile flow and not to a rise in hydraulic conductivity due to pressure pulsations ('stretched pores'). 5. When the water-exchange function of the lung is considered, flow pattern should be taken into account as an entity in its own right in addition to the steady state or the mean component of blood flow.


The secretion of calcium and phosphorus into milk. 1. The time course of appearance of radioactivity in milk was studied following close-arterial infusion of labelled phosphate, Ca or leucine into the mammary artery of lactating goats. Maximum activities were reached at 1.5 hr in all milk fractions including inorganic soluble phosphate, inorganic colloidal phosphate, casein P, soluble Ca, protein-associated Ca and casein. 2. At 0.5 hr, labelling of the soluble and colloidal phosphate fractions was significantly higher than that of the casein P. 3. Recovery of 32P or 47Ca 3 or more hours after infusion into the cistern of the mammary glands was 98% or greater, indicating that the mammary epithelium is virtually impermeable to [32P]phosphate and 47Ca in the milk to blood direction. 4. Ca and P failed to enter milk in excess of the normal secretion rate when the milk was diluted with isosmotic sucrose given by intraductal injection. 5. These data suggest that milk Ca and phosphate in their various forms are secreted, like protein and lactose, by exocytosis of Golgi vesicles. Unless a paracellular pathway is present, as in oxytocin-treated animals, the milk concentrations are maintained by virtue of the impermeability of the mammary epithelium to these substances.


Fever induced in rabbits by intravenous injection of bovine serum albumin. 1. Bovine serum albumin injected into conscious rabbits produced a dose-dependent fever of short latency and duration of at least 2 1/2 hr. 2. The fever was not due to contamination by other proteins nor due to contamination by bacterial endotoxin. 3. The fever following albumin injection had characteristics similar to endogenous pyrogen fever. 4. Some traditional ways of distinguishing bacterial endotoxin fever from other types of fever need to be reassessed.


Electrophysiological and sensory properties of the thalamic reticular neurones related to somatic sensation in rats. 1. In urethane-anaesthetized rats, studies were made on unit discharges of neurones of the thalamic reticular nucleus (tr) receiving the somatosensory input. Such units were found distributed in a part of the tr (s-tr) surrounding the anterior and anterolateral borders of the ventrobasal nuclear complex (vb). 2. A series of five to ten bursts of grouped discharges were elicited in the s-tr neurones as orthodromic responses to single shock stimulation of the medial lemniscus (ml) or the somatosensory cortex (smc). In contrast to this, the vb relay cells were fired by ml impulses only once at short latencies, though later there occurred a series of two to three bursts of grouped discharges. The same response pattern was seen by stimulating the smc, except that the initial response to smc shocks was of antidromic origin. 3. The mean response latency of the s-tr neurones to ml shocks was about 0.6 msec longer than that of the vb relay cells. Ml fibres connected to the s-tr neurones had the same distribution of conduction velocity as those connected to the vb relay cells. It is suggested that most, if not all, of the s-tr neurones are innervated by axon collaterals of the vb relay cells. 4. About half the s-tr units were activated from vibrissae. For most of the remaining half either movements of hairs or touch to the skin were effective. In the vb, the units discharged by movements of hairs were most numerous and those activated from vibrissae were less frequent. While responses of the s-tr neurones to adequate stimuli were almost exclusively of phasic type, 30% of the responses of the vb relay cells were tonic and the remainder were phasic. 5. By exploring the tr extensively, the s-tr was found to be contiguous caudally to the perigeniculate reticular nucleus which is immediately adjacent to the dorsal lateral geniculate nucleus and contains reticular neurones activated by visual stimuli. Though few in number, units were found in the tr which received a convergence of somatosensory and visual inputs or of somatosensory and auditory inputs.


Distribution of climbing fibres on cerebellar Purkinje cells in X-irradiated rats. An electrophysiological study. 1. The distribution of climbing fibres on cerebellar Purkinje cells has been studied with intracellular recordings in X-irradiated and normal rats. 2. In the treated rats, multiple steps in the post-synaptic potential were elicited in 57% of the Purkinje cells by graded stimulation of the climbing fibres, the response was all-or-none in character in the other cells and in all Purkinje cells recorded in normal animals. In the neurones exhibiting the former type of response, no collision was seen along the afferent fibres during interaction experiments between just-threshold juxtafastigial and maximal olivary stimulations, whereas a collision always occurred when all-or-none responses were recorded. 3. These results show that in X-irradiated rats, the majority of Purkinje cells have a multiple innervation by two to four climbing fibres, instead of the one-to-one relationship seen normally. 4. Input resistances and total electrotonic lengths of Purkinje cells were measured in normal and treated rats. Mean values for these two parameters were higher than normal in multiply innervated cells. 5. Mean time course and mean current for reversal of the post-synaptic potential elicited in Purkinje cells by stimulation of the climbing fibres were nearly the same in mono- and in multiply innervated neurones. In multiply innervated cells, time courses and currents for reversal were independent of the size of the response or varied slightly with it, suggesting that the climbing fibres involved innervated territories whose electrotonic distance from the recording site were either the same or slightly different. 6. Interactions between two all-or-none steps of the graded post-synaptic potential evoked in multiply innervated cells by juxtafastigial and olivary stimulations revealed either a very weak or a very marked shunting effect between synapses of the two climbing fibres involved. 7. These results indicate that the over-all distribution of climbing fibre synapses on multiply innervated Purkinje cells is not grossly abnormal and that two fibres contacting a given cell can be either intermingled on the same dendrites, or segregated on distinct dendritic branches. 8. In general, the present study does not suggest the existence of a strong competition among climbing fibres innervating each Purkinje cell during development at least when granule cells are absent.


Immunological detection of somatostatin in a primitive hypothalamic mouse cell line, precursor of a neurophysin cell lineage. A primitive nerve cell line (F7 clone), obtained by SV 40 transformation of mouse fetal hypothalamic cells, has been previously shown to share some properties with a progenitor cell for neurophysin synthesizing neuron. This paper describes the immunological detection, at the light microscope level, of somatostatin in this cell line. A SV 40 transformed neurosecretory clone, synthesizing neurophysin and vasopressin, was used as control.


Hypothalamic neurones in culture. II. A progress report. Cultures of the hypothalamic supraoptic nucleus area were prepared from 1--10 day old rats by means of the roller-tube technique. After 3 weeks in vitro large nerve cells with long, thin and repeatedly branching axons could be observed. In silver-stained co-cultures of hypothalamic and hypophysial tissue, fibers were seen to bridge the gap between the adjacent explants. Intracellular current injection into the large nerve cells elicited single or repetitive action potentials. Two types of spontaneous discharge patterns were observed. The majority of cells fired randomly and the remainder displayed a phasic dicharge pattern. Simultaneous recordings from 2 cells disclosed that pairs of phasic cells had a tendency to fire in synchronous bursts.


[Immunocytochemical study of the maturation of the hypothalamo-neuro-hypophysial axes in the human fetus (author's transl)]. 1. The development of the hypothalamo-neurohypophysial system was studied using immunocytochemistry with various antisera : anti-neurophysin, anti-oxytocin, anti-vasopressin, anti-vasotocin and anti-somatostatin. 2. Immunocytochemical investigation shows that anti-vasopressin and anti-vasotocin sera react with both vasopressin and vasotocin, whereas the anti-oxytocin serum specifically reveals the oxytocin-containing structures (Fig. 1g, h, i). 3. Perikarya stained with anti-neurophysin, anti-vasopressin and anti-vasotocin sera can be seen from the 11th week of fetal life (Fig. 1a) first in the supra-optic nucleus (SON), then in the para-ventricular nucleus (PVN). Their axons reach the neural lobe as early as the 11th week (Fig. 1b, c). 4. Oxytocin-containing perikarya appear simultaneously in the PVN (Fig. 1e) and SON during the 13th week. 5. From the 16th week, neurons stained with the anti-somatostatin serum can be seen among the neurophysin-positive cell-bodies of the SON and PVN. 6. The various populations of magnocellular neurons show a significant increase in number, especially up to the 19th week, and an increase in their hormonal content up to birth.


Relationship between plasma and uterine oestradiol in pseudopregnant rabbits. Plasma oestradiol concentrations did not change during hCG-induced pseudopregnancy except for a slight increase on Day 18. However, a marked decline was observed in tissue oestradiol on Day 3 whether expressed on the basis of tissue wet weight or protein. The lowest concentration was found at Day 6 but the Day 18 values were comparable to those of untreated controls.


The concentration of carnitine in the luminal fluid of the testis and epididymis of the rat and some other mammals. Luminal fluid was collected by micropuncture techniques from the testis and epididymis of the rat, hamster, rabbit, boar and ram and the concentration of free L-carnitine in the fluid was estimated using enzymic methods. Carnitine was present in the testicular fluid of the rat in concentrations less than 1 mM but increased down the epididymis to reach 53 mM in luminal fluid from the cauda epididymidis, approximately 2000 times higher than in blood plasma. A high concentration was first found in the luminal fluid from the distal caput epididymidis, at about the point where the spermatozoa become motile. Carnitine was also present in the epididymal luminal fluid of the other species studied; the amounts were not as high as those in the rat but were still higher than those in blood plasma.


Survival of frozen mouse embryos after rapid thawing from -196 degrees C. The effect of the rate of rewarming on the survival of 8-cell mouse embryos and blastocysts was examined. The samples were slowly cooled (0.3--0.6 degrees C/min) in 1.5 M-DMSO to temperatures between -10 and -80 degrees C before direct transfer to liquid nitrogen (-196 degrees C). Embryos survived rapid thawing (275--500 degrees C/min) only when slow cooling was terminated at relatively high subzero temperatures (-10 to -50 degrees C). The highest levels of survival in vitro of rapidly thawed 8-cell embryos were obtained after transfer to -196 degrees C from -35 and -40 degrees C (72 to 88%) and of rapidly thawed blastocysts after transfer from -25 to -50 degrees C (69 to 74%). By contrast, for embryos to survive slow thawing (8 to 20 degrees C/min) slow cooling to lower subzero temperatures (-60 degrees C and below) was required before transfer to -196 degrees C. The results indicate that embryos transferred to -196 degrees C from high subzero temperatures contain sufficient intracellular ice to damage them during slow warming but to permit survival after rapid warming. Survival of embryos after rapid dilution of DMSO at room temperature was similar to that after slow (stepwise) dilution at 0 degrees C. There was no difference between the viability of rapidly and slowly thawed embryos after transfer to pseudopregnant foster mothers. It is concluded that the behaviour of mammalian embryos subjected to the stresses of freezing and thawing is similar to that of other mammalian cells. A simpler and quicker method for the preservation of mouse embryos is described.


Influence of dose, repeated treatment and batch of hormone on ovarian response in heifers treated with PMSG. The effect of two dose levels (1000 and 2000 i.u.) of three different commercially available batches of PMSG on the ovarian response (ovulations and follicles greater than 10 mn) of 42 heifers was examined in a randomized incomplete block experiment. Each animal was subjected to two consecutive but different treatments. A significant effect of dose was observed and there were fewer ovulations, but no reduction in the number of follicles, after the second PMSG treatment. There was no evidence that the ovarian response was affected by the PMSG batch used.


Proteins in the uterine secretions of the cow. Cows of normal reproductive history were treated with progesterone for periods of 2--3 months. The uterine secretions yielded 5 major fractions of macromolecular components. Two of the fractions comprised serum proteins. In the other 3 fractions at least 9 non-serum proteins were observed, 7 in one fraction, but in only small amounts. Of the remaining 2 non-serum proteins one is an acid phosphatase recoverable in small amounts, and the other is lactoferrin and is the major non-serum protein present in the uterine secretions of progesterone-treated cows.


The role of the seminal vesicles, coagulating glands and prostate glands on the fertility and fecundity of mice. Female mice were allowed to mate with males which had been sham-operated (Group 1); had seminal vesicles, coagulating glands and ventral and dorsal prostate glands removed (Group 2); had the seminal vesicles removed (Group 3); had the coagulating glands removed (Group 4) or had the ventral and dorsal prostate gland removed (Group 5). The pregnancy rate was normal in Groups 1 and 4, severely reduced in Groups 2 and 3 and less so in Group 5. Litter size was reduced in Groups 2 and 3 but not in Group 5. It is suggested that the seminal vesicles and possibly the prostate glands are important in the production of young in mice.


Prostaglandin-induced luteolysis in pregnant and pseudopregnant rabbits and the resultant effects on the myometrial activity. The effect of prostaglandin (PG)-induced luteolysis on the myometrial activity in 20--21-day-pregnant and 11--12-day-pseudopregnant rabbits was studied by intrauterine pressure (IUP) recording during PG infusions. The same dose of PG (10 micrograms/h during 8h) was also given to 7 non-pregnant (untreated) does that were used as controls. Peripheral plasma concentration of progesterone and oestradiol-17 beta were measured at 2-h intervals during the infusion. Plasma progesterone level decreased significantly within 2 h or the start of infusion in pregnant and pseudopregnant does and continued to decrease; at the end of 8 h, the concentrations were 31 and 41%, respectively, of the pre-infusion levels. The amplitude of uterine contractions increased significantly after 4 h in pseudopregnant does, increased slightly but insignificantly in the pregnant does and showed no significant change in the non-pregnant does during PG infusion. The amplitudes developed in the pregnant and pseudopregnant does were significantly different. The direct effect of progesterone (1--3 micrograms/h during 4 h) was also studied in 7 non-pregnant rabbits. After 2 h the amplitude of contractions had decreased markedly and the pattern of activity had become irregular. The results support the concept of a myometrial inhibitory factor other than progesterone in rabbit pregnancy and suggest that this factor(s) originates in conceptus.


Effect of sodium nitrite on the alpha-chlorohydrin-induced lesion of the testis--epididymis complexin the rat. Administration of vasodilator, sodium nitrite (20 mg/kg body weight), 30 min before alpha-chlorohydrin treatment (90 mg/kg body weight) prevented the chlorohydrin-induced lesion in rat testis--epididymis complex. However, administration of vasodilator 90 min after alpha-chlorohydrin treatment did not prevent the chlorohydrin-induced lesion in the testis--epididymis complex. These observations suggest that the testicular vasculature is involved in drug action.


A microsystem for the extraction and in-vitro translation of mouse embryo mRNA. A micromethod is described for the extraction of mRNA from mouse ova and its translation in a message-dependent rabbit reticulocyte lysate system. Using this technique it has been possible, by SDS polyacrylamide gel fluorography, to visualize the products from the equivalent of 12.5 eggs.


Investigation by luminal perfusion of the transfer of compounds into the epididymis of the anaesthetized rat. Controlled perfusion through the lumen of the distal cauda epididymidis in the anaesthetized rat has been explored as a means of examining physiological exchanges from blood across the epididymal epithelium. The mean length of the perfused, sperm-free, tubule was 14.5 cm (+/- 1.5 s.e.m., n = 9). No cholesterol, protein or sialic acid was detected in the perfusate at flow rates exceeding 10 microliters/min, but at rates of 0.4--1.2 microliters/min, protein appeared at concentrations of 0.21--0.55 mg/ml (i.e. secretion rates of 0.21--0.83 micrograms/min; 3 rats). Glucose was detected at all perfusion rates (3--27 microliters/min) at concentrations of 0.06--0.58 mM (0.8--6.8% blood levels). During intravenous infusions of 3H2O, radioactivity in the perfusate rapidly attained 87% blood plasma concentrations; no radioactivity was detected when carboxy-E114C]dextran or methoxy-[3H]inulin were infused. Radioactivity appeared in the epididymal perfusate to 1--7% of blood levels during intravenous infusions of D-E1U-1RC]glucose or 3-O-methyl[1-3H]glucose. This evidence suggests that the preparation is physiological and could be used to explore the dynamics of exchanges between blood and epididymis.


Factors affecting the entry of testosterone into the lumen of the cauda epididymidis of the anaesthetized rat. When [3H]testosterone was infused into the general circulation of the rat, perfusion of a length of the cauda epididymidis (17 +/- 1.0 (s.e.m.) cm, n = 36) with perfusates of varied composition revealed a low entry of radioactivity (1--10% plasma levels; 10 exps) with protein-free perfusates, and a greater entry (15--48%; 10 exps) when the perfusate contained bovine serum albumin (38 mg/ml). When the perfusate contained ovine or rat testicular fluid, or rat epididymal fluid at protein concentrations of 3 mg/ml or less, the entry of radioactivity into the epididymis was greater than when the perfusate contained 3 mg BSA/ml. The addition of ovine rete testis fluid protein (3 mg/ml to BSA (38 mg/ml) in the perfusate increased the uptake of radioactivity (58--106%; 6 exps). Radioactivity in blood was principally associated with testosterone (90, 95% total blood activity, 2 rats), whereas both [3H]testosterone (37, 41% total perfusate activity) and [3H]dihydrotestosterone (42, 63% total perfusate activity) was present in BSA-containing perfusates. The proportion of dihydrotestosterone appeared to increase when the perfusate contained protein of testicular origin.


Steroidogenesis and prostaglandin synthesis by cultured bovine blastocysts. Bovine blastocysts were collected at Days 13, 15 and 16 and placed in TCM-199 supplemented with 5% fetal calf serum; some blastocysts were immediately frozen while the others were cultured for 48 h and then frozen. Samples (tissue + medium, 5--12/group) were thawed, homogenized and analysed by radioimmunoassays. Measurable amounts of progesterone were found in all blastocysts but values were higher (P less than 0.01) after culture. Testosterone was not found in the cultured or uncultured blastocysts at Day 13, but was detectable on Days 15 and 16 and in greater amounts (P less than 0.05) in the cultured blastocysts. PGF and PGE-2 were increased (P less than 0.05) in the cultured blastocysts in all 3 days. Oestradiol was measurable in some but not all blastocysts. It is suggested that PG synthetase and enzymes capable of synthesizing progesterone, testosterone and, possibly, oestradiol are present in these early bovine blastocysts.


Effects of cycloheximide on the uterine refractory state induced by 'nidatory' oestrogen in rats. After priming with oestradiol, ovariectomized rats were given 6 days of progesterone treatment in which two doses of 50 ng oestradiol were given on Days 3 and 6. This basic treatment allows the oestradiol-induced (1st injection) disappearance of uterine sensitivity to decidual stimuli to occur. Cycloheximide could not mimic oestrogen action in the production of the uterine refractory state. However, a high dose (500 micrograms per animal) of this inhibitor given with the first injection of oestradiol allowed the uterus to remain in a neutral state and to respond to decidual induction after the second dose of oestradiol. By delaying the injection of cycloheximide after the first oestrogen treatment, protein synthesis requisite to the occurrence of uterine refractoriness would not take place within 12 h after the 'nidatory' oestrogen injection.


The lack of effect of ejaculate sequence on sex ratio in rabbits. The influence of the order of the ejaculate on the sex ratio was determined by ejaculating 1 of 5 experimental males 20 times over a period of 2 to 8 h. The sex ratio was noted for the offspring of ejaculates 1, 5, 10, 15 and 20 in a total of 32 litters. The sex ratio of these offspring did not deviate signigicantly from 50:50 (P greater than 0.1) for any ejaculate in the sequence.


Reduced prostaglandin levels in the semen of men with very high sperm concentrations. The prostaglandin levels have been measured in a group of men with sperm concentrations greater than 300 X 10(6)/ml and compared with the levels in men with sperm concentrations of 50 to 150 X 10(6)/ml. The distribution of the PG levels in all groups was highly skewed but the data could be transformed to a normal distribution by taking logarithms. Comparison of the PG levels showed a highly significant lowering of the PG levels in the polyzoospermic group when compared wieth either of the groups with normal sperm concentrations.


Effects of fluoroacetate on the testis of the rat. Rats recieving 20, 6.6 or 2.2 p.p.m. sodium fluoroacetate in the drinking water were killed daily during the 7 days of treatment and at more widely spaced intervals in the succeeding 21 days. Testicular weight and ATP concentrations decreased in rats receiving 20 or 6 p.p.m. fluoroacetate, while citrate concentrations were elevated and morphological damage was seen in the testes of all the treated rats. Initial cellular changes common to the three treatment groups included altered appearance and decreased numbers of spermatids, and formation of spermatid and spermatocyte giant cells. At the two higher concentrations damage progressed to marked seminiferous tubule atrophy. Regeneration of the seminiferous tubules was complete by 7 days after treatment, in the rats given 2 p.p.m. but regeneration was not complete by Day 21 after treatment in those receiving the higher doses. Spermatogenesis was abnormal in some instances during the regneration period in these groups. The findings are consistent with impaired energy production via blockage of the Krebs cycle, and subsequent impairment of carbohydrate metabolism through the Embden-Meyerhof pathway.


A cytogenetic survey of men being investigated for subfertility. A study of 336 men attending a subfertility clinic but otherwise unselected, and a further 12 men studied as a part of the investigation of the female partner, revealed 10 with major chromosome anomaly and 3 with unambiguous chromosomal variants. In addition to those with sex chromosome aneuploidy, an extra, small marker chromosome and D/D Robertsonian translocations, anomalies which have been reported in other studies, there were 7 men with rearrangements, including a paracentric inversion of chromosome 7 and an X/21 reciprocal translocation. These would have been difficult or impossible to identify without good banded preparations, suggesting that such rearrangement may be more frequent in association with subfertility than was appreciated.


Androgen receptors in the rat epididymis and their hormonal control. The concentrations of cytoplasmic receptor sites for androgens in the caput, corpus and cauda epididymidis, and the effect of ligation of the efferent ducts and testosterone treatment after bilateral castration on the concentration of receptors in the caput have been measured. Androgen receptors in the ventral prostate have been measured in the same animals for comparison. The caput has the highest concentration of receptor sites, the corpus the lowest. The ligation of the efferent ducts has no effect on this concentration which is dependent on testicular androgens. The present data do not yet allow explanation of the differential response of the different regions of the epididymis and of the other accessory glands to the administration of androgens.


Amino acid transport in the unfertilized and fertilized mouse egg. The transport of methionine into unfertilized and fertilized mouse eggs appears to involve active transport mechanisms with similar Vmax, Km, substrate specificity and independence from Na+. An exchange diffusion system with a similar amino acid specificity to the uptake system has also been found in both types of egg. An estimate of 6.5 fmol has been made for the size of the total internal pool of exchangeable amino acids.


The effects of cooling to 5 degrees C and freezing and thawing on the ultrastructure of bull spermatozoa. Semen from 6 bulls was examined under the transmission electron microscope immediately after collection, after dilution and cooling to 5 degrees C and after freezing and thawing. Conception rates were determined following artificial insemination of the frozen and thawed semen. Dilution and cooling to 5 degrees C caused acrosomal swelling in about 50% of the spermatozoa. Subsequent freezing and thawing caused considerable ultrastructural changes to the acrosomes (disruption of the plasma and outer acrosomal membranes and dispersion of the acrosomal contents) and middle pieces (breakage of the plasma membrane and a reduction in the electron density of the mitochondrial matrix) of a high proportion of spermatozoa. The average non-return rate following insemination of semen from 5 of the bulls was 61.6% and higher (P greater than 0.001) than for the sixth bull (15%). Although this difference in semen viability was also demonstrated in the structural studies (acrosome, P greater than 0.05: middle piece, P greater than 0.001), more work is required to assess the relationship between structure and function of spermatozoa.


Two intersex dogs with mosaicism.

Two intersex dogs with mosaicism. Two intersex dogs with various degrees of mosaicism in the somatic tissues are described. Normal female cells as well as Y-bearing cells with aneuploidy and other abnormalities were involved. In the Red setter an ovarian cortex had developed in mosaic gonads with 8 and 10% of Y-bearing cells. Slight masculinization of the medullae corresponded with some external virilization. These observations support the existence of a threshold proportion of Y-bearing cells for testicular differentiation. Fragments of chromatin in 36% or fewer of the cells in the Cocker spaniel may have been translocated Y-chromosome material.


The effects of corpus luteum removal during gestation on parturition in the tammar wallaby (Macorpus eugenii). Removal of the corpus luteum of pregnancy of the tammar wallaby interfered with successful parturition if carried out before Day 17 of the 27-day pregnancy. After removal at Days 17 and 21, 40% of animals gave birth but pouch young died with 24 h; if performed at Days 23 or 25, pouch young survived. However, surgery also affected sham-operated controls if performed between Days 15 and 21. In control animals, sodium pentobarbitone followed by halothane anaesthesia was the least disruptive anaesthetic procedure. This study shows that the corpus luteum has an essential role in parturition and subsequent survival of the neonate in the pouch.


Synthesis of 'decidualization-associated protein' in tissues of the rat uterus and placenta during pregnancy. The synthesis of a soluble protein (referred to as 'decidualization-associated protein', DAP), has been examined in uterine and placental tissues of rats during pregnancy by means of polyacrylamide gel electrophoretic analysis of [3H]leucine-labelled soluble proteins. No synthesis of the protein was detected in non-implantation regions of the uterus. In implantation site tissue, no synthesis was detected on Days 6 or 7 of pregnancy. Only slight synthesis was present in the endometrium on Day 8, but synthesis rose rapidly from Days 9 to 12 in both the endometrium and myometrium although differences in the rates of increase were observed. Synthesis fell from Day 12 to 14 in both tissues. Synthesis by the myometrium was entirely localized in the mesometrial region, which contains the metrial gland. After Day 12, when the endometrium is represented by the chorioallantoic placenta, synthesis was examined in the labyrinthine and the decidua basalis/basal zone placenta tissues. No synthesis of 'DAP' was detected in the labyrinthine placenta from Day 16 of pregnancy. Synthesis observed in the decidua basalis/basal zone placenta fell dramatically from Day 14 to 20. The pattern of synthesis of 'DAP' during pregnancy suggests a role in the establishment of the chorioallantoic placenta and metrial gland in the rat.


Comparison of the embryo survival rate in heifers following artificial insemination, non-surgical blastocyst transfer or both. In Exp. 1, embryo survival rates of 45 and 47% were recorded after artificial insemination and ipsilateral transfer respectively. In Exp. 2, pregnancy rates of 62 and 60% were recorded after artificial insemination and contralateral transfer to inseminated recipients respectively. In this experiment the contralateral transferred embryo survival rate was 44%. Transferred embryo survival was lower overall when donors and recipients were out of phase by 1 day than when exactly synchronous.


Changes in uterine phosphatase levels in mice deprived of LH during early pregnancy. Mice injected with normal rabbit serum on Day 4 of gestation showed a progressive increase in specific alkaline and acid phosphatase activities on Days 6 and 7. Mice injected with antiserum to LH on Day 4 showed a significant decrease in specific alkaline phosphatase activity estimated biochemically on Days 6, 7 and 8, but this decrease was histochemically evident only on Day 8. Similarly, acid phosphatase activity in antiserum-treated mice was significantly increased on Days 6, 7 and 8 when estimated biochemically, but only on Day 8 when studied histochemically.


Early pregnancy diagnosis in the ewe, based on milk progesterone levels. Plasma and milk progesterone concentrations in pregnant sheep (18--22 days after mating) were similar, about 3.7 ng/ml whereas values in non-pregnant sheep were less than 1 ng/ml. Lambing results indicated identical accuracy for both methods (82 and 84% in 2 flocks). The accuracy was 92--100% for ewes diagnosed non-pregnant in the breeding season, but for ewes tested in the non-breeding season the diagnosis of non-pregnancy according to milk progesterone levels was only 50% accurate.


Presence of placental lactogen in bovine conceptuses before attachment. Placental lactogen has been detected by radioreceptor assay in bovine conceptuses colletected between 17 and 25 days post coitum, at or shortly after the time of appearance of binucleate cells in the bovine trophectoderm, but before attachment or implantation.


Preliminary observations on the effects of 19-hydroxy prostaglandin E-1 on the activity of the human myometrium in vitro. The principal prostaglandin of human semen, 19 hydroxy PGE-1, inhibited spontaneous contractile activity of myometrium from non-pregnant women in vitro. This inhibition was similar to that caused by PGE-1. The inhibitory effects of natural 19 hydroxy PGE-1, prepared from semen of Macaca arctoides, were similar to those of synthetic 19 hydroxy PGE-1.


Steroid receptors in the canine epididymis. High-affinity and low-capacity molecules that bind 5 alpha-dihydrotestosterone and oestradiol-17 beta have been demonstrated in the canine epididymis. The molecules were shown to be proteinaceous in nature, thermolabile and sedimented in a sucrose density gradient with a coefficient of 8S with respect to bovine serum albumin. Only minor differences were found for dissociation constants for oestradio-17 beta (3.5 +/- 1.74 S 10(-10) M) and 5 alpha-dihydrotestosterone (4.2 +/- 10.75 X 10(-10) M) (means +/- s.d.) in the three anatomically distinct regions of the epididymis.


Uterine blood flow of cows during the oestrous cycle and early pregnancy: effect of the conceptus on the uterine blood supply. Blood flow to each uterine horn of cows during the oestrous cycle and early pregnancy was determined daily by use of electromagnetic blood flow probes placed around both middle uterine arteries. The pattern of blood flow to uteri of pregnant and non-pregnant cows was similar until Day 14 after mating or oestrus. Between Days 14 and 18 of pregnancy blood flow to the uterine horn containing the conceptus increased (P less than 0.01) 2- to 3-fold, whereas blood flow to the non-gravid uterine horn in these cows remained constant. No corresponding increase in blood flow to the uterine horn ipsilateral to the ovary bearing the CL was observed in non-pregnant cows during this 4-day period. By Day 19 of pregnancy, blood flow to the gravid uterine horn had returned to a level similar to that observed on Day 13. Blood flow to both uterine horns of pregnant cows remained constant from Days 19 to 25 and then increased to the gravid horn (P less than 0.01) markedly until Day 30 whereas blood flow to the non-gravid horn remained low. Uterine blood flow during the oestrous cycle of non-pregnant cows was positively correlated (P less than 0.01) with systemic concentrations of oestradiol and the ratio of oestradiol (pg/ml) to progesterone (ng/ml). There was no association between oestradiol concentrations and blood flow to the gravid uterine horn. These data indicate local control of uterine blood flow by the bovine conceptus which may function to create optimal conditions for the continuation of pregnancy.


Peripheral plasma androstenedione and testosterone concentrations in bulls before and during puberty. Peripheral plasma samples were collected from 37 young bulls every other month from 2 to 12 months of age. Androstenedione and testosterone were measured by radioimmunoassays. Androstenedione values increased from 2 to 4 months of age (525 +/- 296 (s.d.) pg/ml) and then decreased. Testosterone concentrations increased regularly from 2 months (0.17 +/- 0.14 ng/ml) to 6 months (2.79 +/- 1.29 ng/ml) and then remained at levels around 3 ng/ml. Coefficients of correlation between the hormone concentrations were significant only at 6 months and thereafter.


Trophoblastin, an antiluteolytic protein present in early pregnancy in sheep. Trophoblastin, an antiluteolytic component from the embryo, was identified in the ewe by the means of intrauterine injections of homogenates from trophoblasts at 14--16 days pregnancy. Homogenates from embryos and their membranes at 21--23 days pregnancy did not extend the life of the corpus luteum, suggesting that trophoblastin synthesis occurs for only a short period. The trophoblastin was thermolabile (80 degrees C for 30 min) and inactivated by pronase. Treatment of ewes with oCS, hCG, and extracts of 120-day placentae did not affect the time of luteolysis. The protein appears to be insoluble at pH 7 or 8, but to dissolve readily at pH 9.6. After injection of homogenates or extracts from 15--16-day-old trophoblasts, the initial CL were maintained for more than 1 month in most cyclic recipient ewes. Surgical removal of embryos at 21--23 days resulted in luteal maintenace for more than 1 month in over 50% of the operated animals. All the maintained CL were secretory although their average weight was about one-half of that CL of normal pregnancy, suggesting the existence of complementary luteotrophic placental factors. The uteri of most of these pseudopregnant ewes were distended with a clear, sterile fluid.


Detection of early pregnancy in sheep by the rosette inhibition test. The rossette inhibition test, an established test for determining the immunosuppressive potential of antilymphocyte serum, has been applied to the serum of sheep after mating. The rosette inhibition titre was much higher (12--26) in 7 sheep which were fertilized and remained pregnant for up to 21 days than in 5 sterile ewes mated with intact rams (8--10). The difference was apparent by 24 h after mating. One ewe had high titres for 6 days after mating but these then dropped and she returned to oestrus; early embryonic loss was suspected. Another ewe which returned to oestrus had consistently low titres. The results indicate that the rosette inhibition test can be used to detect fertilization, early embryonic death and continued pregnancy in sheep.


Effect of prostaglandin F-2 alpha on plasma levels of progesterone and pregnenolone in the hysterectomized guinea-pig. Repeated injections of PGF-2 alpha were given to hysterectomized guinea-pigs. Within 12 h after the first injection, plasma progesterone levels were significantly reduced (P less than 0.001) and declined to less than 1 ng/ml by the 4th day after the end of PGF-2 alpha treatment. A decline in plasma levels of pregnenolone paralleled that of progesterone. PGF-2 alpha treatment did not affect the metabolic clearance rate of [3H]pregnenolone, but the converions of [3H]pregnenolone to [3H]progesterone was reduced by about 50%.


Effect of prostaglandin F-2 alpha on ovarian enzyme activity in the hysterectomized guinea-pig. The enzymes studied were cholesterol esterase, cholesterol ester synthetase 3 beta-hydroxysteroid dehydrogenase and 20 alpha-hydroxysteroid dehydrogenase. PGF-2 alpha reduced the activities of 3 beta-hydroxysteroid dehydrogenase and cholesterol esterase but did not affect those of cholesterol ester synthetase of 20 alpha-hydroxysteroid dehydrogenase.


Luteinization of bovine granulosa cells and corpus luteum formation associated with loss of androgen-aromatizing ability. The relative aromatizing ability of bovine luteinizing granulosa cells and dispersed luteal cells in tissue culture was studied. Luteinization of granulosa cells, as indicated by steadily increasing progesterone production (from 50 to 300 ng/10(5) cells/day over 4--5 days), was accompanied by a dramatic reduction in their capacity to aromatize exogenous androgen; oestradiol-17 beta production falling from 200 to less than 10 ng/10(5) cells/day over 4--5 days. Luteal cells also had only a very limited capacity to aromatize exogenous androgen, maximum oestradiol-17 beta production being less than 600 pg/10(5) cells/day. The loss in aromatizing capacity of granulosa cells during luteinization was also reflected in the relative endogenous steroid content of non-luteinized granulosa cells and luteal tissue, the former containing high levels of oestradiol-17 beta, less than or equal to 28 ng/mg protein, while the latter, although containing substantial amounts of testosterone, less than or equal to 5.7 ng/g tissue, contained very little oestradiol-17 beta, less than or equal to 0.35 ngG TISSUE. These findings suggest that luteinization of bovine granulosa cells and subsequent corpus luteum formation is associated with a loss in androgen aromatase activity.


Fibrocystic disease of the breast.

Fibrocystic disease of the breast. Fibrocystic disease is the most common type of lesion in the female breast. It is a common dumping ground for a variety of distinct clinical and histologic entities which require different modalities of therapy and which have different malignant transformation potentials. In general, fibrocystic disease is managed medically unless a dominant lump or other adverse symptom develops, in which case a biopsy is performed. The exception to this rule is a cyst which can be safely aspirated under controlled conditions. If, on biopsy, one of the histologic types of fibrocystic disease called "precancerous mastopathy" is found, which has a high potential for malignant transformation, then serious consideration must be given to definitive surgical procedures, especially if the patient has other risk factors that would increase her risk of developing breast cancer even further.


Law and medical ethics.

Law and medical ethics. Summarising the interrelationship between law and medical ethics, I would say that in cases which do not touch the patient's body or integrity, such as professional secrecy, statutory law may take precedence over rules of medical ethics. But in cases where the human subject becomes a victim because of domestic statutory laws which are in contradiction with medical ethics, the medical practitioners should insist on adhering to their professional standards in such a way that the legislators will have to adapt their legislations to the laws of humanity and public conscience. Legislators, as well as medical practitioners, should not forget that the term 'being' is preceded and qualified by 'human'.


The nature of confidentiality.

The nature of confidentiality. This paper examines confidentiality and its nature and analyses the guidelines laid down by the Hippocratic Oath as well as the British and World Medical Associations for maintaining such confidentiality between doctor and patient. There are exceptions to practically any code of rules and this is true also for confidentiality. Some of these exceptions make it appear that very little is confidential. The three values implicit in confidentiality would seem to be privacy, confidence and secrecy. Each of these values is discussed and developed in this paper. In conclusion, the question is suggested that maybe in the face of death, doctor and patient need to re-examine the pre-suppositions of privacy, confidence and secrecy on which the confidential relationship is based.


Deciding the care of severely malformed or dying infants. Suffering patients (when able), grieving families and compassionate physicians have always sought the least detrimental alternative while deciding care in the face of tragedy. Modern medical technology has brought great benefits to patients but has blurred traditional concepts of life and death and created new dilemmas for practising doctors. While this technology has given doctors great control over living and dying, their dominance in critical decision making is being challenged. More and more their decisions are liable to public and legal scrutiny, intense publicity by the news media and exploitation by lobbyists with opposing aims. Increasing pressure of this kind may deflect the physician form his primary responsibility to patients and their families. For infants with gross malformations or a distressing terminal illness we believe that the parents and their doctors must be allowed primary decisional power even if the chosen course of action involves the death of the infant. Choices for death should be permitted but only after suitable family and professional consultation. Some general guidelines are suggested. As these situations are so varied and so complex, much latitude in decision-making should be expected and tolerated.


Learning about death.

Learning about death. This paper outlines briefly some of the research which has been carried out on attitudes to terminal illness and its care. The writer feels that not enough effort is being put into the teaching of this subject in our medical schools and Universities, and that doctors themselves are the ones who often wish to 'duck' the issue of dealing with disability and the dying. However, with the increasing awareness, through both the research and the growing allied literature, the writer feels that there is no longer any excuse for omitting this subject from the curricula for doctors and nurses in training.


Familial caudal regression anomalad and maternal diabetes. A family is reported which a diabetic woman gave birth to two children with the caudal regression anomalad (CRA). There were no obvious genetic factors. This is the first reported familial case of CRA with maternal diabetes.


Tissue typing amniotic fluid cells: potential use for detection of contaminating maternal cells. The presence of contaminating maternal cells in amniotic fluid is an important, though infrequent, cause of error in karyotyping the fetus. A method of detecting contaminating maternal cells in amniocentesis specimens by determining the HLA phenotype of the cells of amniotic fluid and the mother is described. Tissue typing of 15 amniocentesis specimens was performed, and in 14 cases the fetal origin of the cells was established. In one case, the results of tissue typing suggested maternal cell contamination, though this had not been suspected from chromosome studies of the amniotic fluid cell cultures. Other possible uses for tissue typing of amniotic fluid specimens for prenatal diagnosis are also described.


Tuberous sclerosis: possible modification of phenotypic expression by an unlinked dominant gene. A unique pedigree is presented which shows tuberous sclerosis in three generations of a family, in which two heterozygotes for the mutant gene were found to be clinically asymptomatic. A genetic model is proposed to explain these findings based upon the segregation of a second unlinked autosomal dominant gene modifying the expression of the gene for tuberous sclerosis.


Serum creatine kinase levels in normal females. Serum creatine kinase levels were determined in 75 girls (age range, one month to 15 years) and 200 normal adult women (age range, 18 to 50 years). The values ranged from 12.5 to 80 IU/1 in girls and 19 to 155 IU/1 in adult females. The SCK level appeared to increase with age from 1 to 15 years, after which the level remained fairly constant. These data should be helpful in the detection of carriers of X-linked forms of muscular dystrophy.


Beta-thalassaemia: increased chromosomal anomalies in lymphocyte cultures. Lymphocyte cultures from homozygotes for the beta-thalassaemia gene were found to contain chromosomal gaps, breaks, and rearrangements more often than those from controls. Culture time seemed to have a determinant effect on the results. The possible influence of folic acid deficiency is discussed.


Meiosis in two human reciprocal translocations. Analysis of meiotic chromosomes from two human male translocation heterozygotes is reported. One had total sterility after maturation arrest and chain configurations at meiosis, and the second had a abnormal children who were presumed to be genetically unbalanced.


Familial partial 14 trisomy.

Familial partial 14 trisomy. Four children in the same family have 47, +der (14), t(9;14) (p24;q21). Their mothers are sisters with 46,XX,t(9;14) (p24;q21). Clinical features of the children are similar to those of others reported to have partial 14 trisomy.


Extra small metacentric chromosome identified as i(18p). A case of a supernumerary metacentric small chromosome, diagnosed at birth, is described. The cytogenetic findings support its identification as i(18p). The clinical development from birth to 12 months is reported, with particular attention given to the psychomotor retardation and to the immunological aspect.


Familial dicentric translocation t(13;18)(p13;p11.2) ascertained by recurrent miscarriages. A dicentric translocation is described involving chromosomes 13 and 18 in which the centromere of chromosome 13 was suppressed. The translocation was ascertained by repeated miscarriages and was found in three generations of phenotypically normal carriers.


The 9p-syndrome.

The 9p-syndrome. A 13-year-old boy with 9p- (p22 leads to pter) is reported. He had many features in common with previous 9p- cases, as well as several distinctive features including polydactyly and precocious puberty. Cytogenetic studies revealed a de novo deletion distal to ban 9p22, which was the reported site of chromosome break in 9 of the 10 previous 9p- cases. Evaluation of the human GALT enzyme suggests that its locus is not on the deleted segment.


Variation in chromosome 19.

Variation in chromosome 19. Variations in centromeric staining of chromosome 19 appear to be an uncommon polymorphism inherited in a Mendelian manner and easily seen in G-banded cells. It should not be misinterpreted as a structural cytogenetic abnormality.