Biomedical subjects
Publications and source records attributed to R Bachvarova.
A method is described for in vitro culture of naked growing mouse oocytes for at least four days. Tests for metabolic function indicate that the oocytes increase in volume, accumulate glucose-6-phosphate dehydrogenase, and maintain a steady level of 3H-uridine and 3H-leucine incorporation.
The RNA of growing mouse oocytes and ovulated ova was labeled by injection of a solution containing tritiated uridine into the ovarian bursa. The time course of incorporation into RNA by oocytes was followed by analysis of alkali-labile acid insoluble radioactivity and by autoradiography. The results show that most of the incorporation into RNA by growing oocytes takes place within one day of bursal injection of the precursor, reflecting the rapid fall of label in the acid soluble precursor pool. The RNA of growing oocytes of all sizes is unusually stable, at least 80% of the labeled RNA present two days after bursal injection being retained until ovulation 10 to 20 days later. The fraction of heterogeneous RNA in labeled RNA of ova was estimated as 20% by sucrose gradient analysis. It is likely that egg RNA is synthesized primarily during the period of oocyte growth one to three weeks before ovulation.
RNA synthesis, previously shown to take place during oocyte growth, has been demonstrated throughout the growth-quiescent period preceding ovulation of the mouse oocyte. In the final 7-day preovulatory period, the level of incorporation of (5,6-3H)uridine into ovulated oocytes decreased as the interval between exposure to precursor and ovulation decreased; significant incorporation was detectable within 2 days before ovulation. Analysis of the frequency and density of label in ovarian oocytes at successive stages of meiosis in relation to the interval between adminstration of labeled precursor and collection of oocytes revealed that RNA synthesis continues up to within 2 h before GVBD.
Mouse embryos exposed to concentrations of 5-bromodeoxyuridine (BUdR) ranging from 0-01 to 1-0 mug/ml in vitro for two days from the 8-cell stage exhibit a concentration-dependent decrease in the frequency of normal blastocysts and decrease in average cell number per embryo. A 20-h exposure was adequate to achieve the full BUdR response. Both effects were eliminated in the presence of excess thymidine. Autoradiographs demonstrated that BUdR[3H] was incorporated into DNA during the first and second day of culture. Thus, BUdR appears to act through incorporation into DNA; and, in this system, cell division is at least as sensitive to BUdR as is differentiation.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Explore the source record for details and available documents.