Mouse dynein axonemal intermediate chain 2: cloning and expression
Ovarian follicular development is a complex process. Investigation of the mechanisms regulating the initiation of follicular growth, and the growth and differentiation of preantral follicles is of great interest. In an effort to clone follicular development-related genes, we selected a partial cDNA fragment by differential display reverse-transcription PCR using total RNA extracted from 5-day-old and 10-day-old mouse ovaries, and its open reading frame was obtained by rapid amplification of cDNA ends. Sequencing showed that the fragment is the mouse dynein axonemal intermediate chain 2 gene (Dnaic2), which has an 87% homology with human DNAI2, a candidate gene for primary ciliary dyskinesia. Northern and western analyses indicate that Dnaic2 produces an approximate 3 kb mRNA that is translated into an approximate 70 kDa protein. The mRNA is predominantly expressed in mouse ovary, testis, and lung. In mouse ovaries, Dnaic2 mRNA was detected at high levels in vivo on day 10, with a subsequent decrease on days 15 and 20, in adult and old ovaries. However, Dnaic2 expression was weak on day 5. Dnaic2 protein was localized on the surface of the oocyte. No obvious fluorescence signal was detected in primordial and primary follicles, while strong signals were detected on the oocyte surface of secondary and antral follicles, in particular for secondary follicles in day 10. These data suggest that Dnaic2 plays a role in ovarian follicular development.