Biochemical studies of mammalian oogenesis: Kinetics of accumulation of total and poly(A)‐containing RNA during growth of the mouse oocyte

Abstract

Kinetics of accumulation of total and poly(A)‐containing RNA have been measured during growth of the mouse oocyte. Total RNA from oocytes isolated at discrete stages of growth was determined by two independent micro‐assays. The fully‐grown oocyte contained about 0.60 ng of RNA. Kinetics of accumulation of total RNA with respect to oocyte volume were biphasic. Small, growing oocytes (about 30 pi) contained about 0.20 ng of RNA/oocyte. The amount of RNA increased in a quasi‐linear fashion until oocyte volume was about 160 pl, at which point there was about 0.57 ng of RNA/oocyte. Thus oocytes about 65% of their final volume had accumulated about 95% of the total amount of RNA present in the fully‐grown oocyte. The relative amount of poly(A)‐containing RNA in oocytes of various size was determined by in situ hybridization of [³H]poly(U) to ovarian sections from juvenile mice of known age, followed by autoradiography. The kinetics of accumulation of poly(A)‐containing RNA were similar to those of total RNA; oocytes about 70% of their final volume had accumulated about 95% of the amount of poly(A)‐containing RNA present in the fully‐grown oocyte. The poly(A)‐containing RNA resided predominantly in the cytoplasm and no obvious cytoplasmic localization was observed. Kinetics of accumulation of total RNA, which is mainly ribosomal, and poly(A)‐containing RNA were consistent with levels of RNA polymerases I and II measured by others during oocyte growth (Moore and Lintern‐Moore, '78). The number of ribosomes that could be made from the amount of rRNA present at various stages of growth was compared to the actual number of ribosomes calculated from a published morphometric study (Garcia et al., '79). Kinetic differences in accumulation between the theoretical and actual number of ribosomes suggested oocyte ribosomes are recruited into cytoplasmic lattice structures. These structures accumulate during oocyte growth and have been postulated to be a ribosomal storage form. In addition, the results from this study are compared to results derived from lower species.