Maturation and ovulation of Japanese quail oocytes under in vitro conditions.

1. An in vitro system for ovulation and maturation of Japanese quail oocytes is described. 2. Ovarian follicles removed from the ovary at 2, 4 or 6 h before the estimated time of ovulation may ovulate under in vitro conditions. 3. The presence of progesterone in the medium had a stimulatory effect on the process of maturation, as has been shown for Xenopus oocytes.

Characteristics of poly(A)-degrading factor present in the avian oocytes and early embryos.

The presence of poly(A)-degrading activity was studied in vitro in the quail and mouse oocytes and early embryos using 3H-poly(A) as a substrate. The activity was measured by adsorption of the undegraded substrate to DE-81 filter paper discs, by chromatographic separation on Sephadex G-50 column and by agarose gel electrophoresis followed by transfer onto a Zeta-probe membrane (BioRad, Richmond, CA) and autoradiography. High poly(A)-degrading activity was found in the quail previtellogenic and vitellogenic oocytes and lower activity in the early embryos from cleavage stage to gastrulation. This activity is localized predominantly in the nucleus and, to a lesser degree, in the cytoplasm and in the vitellus of vitellogenic oocytes. The length of the poly(A) degradation product was estimated to be of about (A)10. Optimum activity was at pH 8.7 and at Mn2+ concentration of 0.5 mM. This makes the deadenylating enzyme from the quail oocytes similar to endoribonuclease IV from the chick and quail oviducts (Müller [1976] Eur. J. Biochem., 70:241-248; Müller [1976], Eur. J. Biochem., 70:249-258). We suggest that the poly(A)-degrading enzyme, similar to endoribonuclease IV found in the quail oocytes, might be the "deadenylating factor" reported in Xenopus oocytes (Varnum et al. [1992] Dev. Biol., 153:283-290). Such poly(A)-degrading activity is undetectable in unfertilized mouse eggs; however, a slight, statistically insignificant tendency for poly(A) degradation was seen in two-cell embryos.

RNase A activity in Japanese quail oocytes.

The presence of RNase A activity was studied in vitro in homogenates of quail oocytes and early embryos using [3H]poly(U) as a substrate. The activity was measured by adsorption of the undegraded substrate onto DE-81 filter paper discs and by chromatographic separation on a Sephadex G-50 column. RNase A activity examined by these methods was almost undetectable in quail previtellogenic, vitellogenic and ovulated oocytes as well as in the embryos from laid eggs. It is estimated to be about 1.1 x 10(-5) Kunitz units per ovulated oocyte. Higher activity starts to appear in gastrulating embryos. These findings are discussed in relation to other studies demonstrating the high stability of maternal RNA during early development, especially in growing oocytes.

Influence of embryonic and adult testis on the differentiation of embryonic ovary in the mouse.

The development of 11 1/2-13 1/2-day embryonic mouse ovaries subjected to the influence of adult and embryonic testes was investigated. The environment of adult testis caused severe restriction of ovarian growth, but did not produce any effects which might be considered as masculinization. The presence of embryonic testis was distinctly unfavourable to the embryonic ovary, resulting in restriction of the growth of the latter and degeneration of oocytes. Reversal of the course of differentiation of genetically female germ cells has never been observed.