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Biomedical subjects

J Brachet

Publications and source records attributed to J Brachet.

At least 37 records · Page 2Linked to original sources

Ionic requirements for induction of maturation (meiosis) in full-grown and medium-sized Xenopus laevis oocytes.

There is already good evidence that calcium ions are involved in the induction of maturation in full-grown amphibian oocytes; we show here that other cations (K+, Mg2+) also play a role in this process. Full-grown (1.3 mm in diameter) and medium-sized (0.8-1.0 mm in diameter) oocytes were compared in the present study. It was found that, provided the medium is K+-free, valinomycin induces maturation in full-grown, but not in medium-sized, oocytes. Increasing the CaCl2 (20 mM) or the MgSO4 (40 mM) content of the medium induces maturation in full-grown, but not in medium-sized, oocytes; however, the latter undergo germinal vesicle breakdown after treatment with either progesterone or ionophore A23187 if there is an excess of Ca2+ or Mg2+ in the medium. Maturation is possible in a Na+-free medium, but amiloride inhibits germinal vesicle breakdown when NaCl is present in the medium. It is concluded that maturation is controlled by changes in the balance between the various ions rather than by Ca2+ alone.

Amiloride↗

[Induction of maturation (meiosis) in small oocytes of Xenopus. A cytological study].

Injection of endoplasm from large Xenopus oocytes which have undergone maturation into small (0.6-0.8 mm diameter) oocytes induces germinal vesicle (GV) breakdown. This process has been cytologically studied. Injection of cytoplasm from such small oocytes into large oocytes (1.0-1.2 mm diameter) is followed by the formation of a large spindle bearing many chromosomes at the site of injection.

Animals↗

Induction of maturation (meiosis) in small Xenopus laevis oocytes by injection of maturation promoting factor.

Treatment of small-(stage III) or medium-sized (stage IV) Xenopus laevis oocytes with progesterone, human chorionic gonadotropin, or para-hydroxymercuriphenylsulfonate does not induce maturation. Only the full-grown oocytes (stage VI) undergo maturation when treated with either one of these three substances. In contrast, injection of maturation promoting factor into oocytes of stages III-VI invariably leads to chromosome condensation and germinal vesicle breakdown. No maturation spindle is found in oocytes smaller than 0.9 nm in diameter, and the nuclear sap does not mix with the cytoplasm in the smallest (0.45-0.55 mm in diameter) oocytes. In oocytes of 0.9 mm, maturation is identical to that of full-grown oocytes, except that the maturation spindle does not reach the cortex of the oocyte. Progesterone increases protein synthesis in medium-sized (0.8 mm in diameter) oocytes without inducing meiosis. It has little or no effect on protein synthesis in smaller oocytes.

Animals↗

[Meiosis: role of a histone kinase in the condensation of ovarian oocyte chromosomes of Xenopus laevis and Ambystoma mexicanum].

By injecting heterologous histone-kinase preparations into ovarian Axolotl oocytes, it has been possible to speed up the progesterone-induced process of chromosome condensation. Moreover, in some instances, this condensation and even complete maturation have been obtained after injection of protein kinase alone, thus in the absence of hormone stimulation. Two different histone kinase preparations have been used: one was prepared from ascites cell chromatin and the other from in vitro ovulated Xenopus oocytes.

Ambystoma↗

Studies on maturation in Xenopus laevis oocytes. III. Energy production and requirements for protein synthesis.

The following results were obtained: 1) The oxygen comsumption of progesterone-stimulated X. laevis oocytes increases at the time of germinal vesicle (GV) breakdown. 2) Continous treatment with 1 mM KCN, 1 mM and 0.1 mM DNP completely inhibits GV breakdown. 3) Pretreatment experiments with KCN, DNP and cycloheximide show that binding of progesterone to the plasma membrane and the specific hormone receptor requires neither energy, nor protein synthesis. 4) 1 h pulses of DNP (1mM) or cycloheximide (50 mug/ml) were applied, at various time intervals after progesterone treatment: early pulses strongly delay, but do not prevent GV breakdown; late pulses just before GV breakdown induce a number of cytological abnormalities, which ultimately lead to cytolysis. The significance of these results is discussed and the hypothesis that synthesis of the maturation producing factor (MPF) requires both energy and protein synthesis is proposed.

Animals↗

Ribonucleotide reductase activity during amphibian development.

Uniformly labeled [3H] uridine is incorporated into DNA by dissociated Pleurodeles blastulae; the label is found in cytosine and to a much lesser extent in thymine. Ribonucleotide reductase activity cannot be detected in full grown oocytes of Xenopus and Pleurodeles, but is present in unfertilized egg. The enzyme is synthesized (or activated) when maturation is induced in Xenopus oocytes by in vitro hormonal treatment. The enzymatic activity increases after fertilization and reaches a peak at the 2--4 cell stage; it decreases at the blastula, gastrula and neurula stages to the low level initially present in unfertilized eggs. The enzyme is no longer detectable in swimming tadpoles. Addition of hydroxyurea (1 mg/ml) to fertilized eggs leads to complete loss of ribonucleotide reductase activity: cycloheximide (20 mug/ml) inhibits the rise in activity characteristic of early cleavage, while actinomycin D (20 mug/ml) has no effect. The significance of these results in discussed.

Amphibians↗

Induction of maturation (meiosis) in Xenopus laevis oocytes by three organomercurials.

Three organomercurials, p-hydroxymercuribenzoate, p-hydroxymercuriphenylsulfonate, and mersalyl, induce maturation (meiosis) in a large percentage (20-100 percent) of Xenopus laevis oocytes. Maturation takes place even when the follicle cells which surround the oocytes have been withdrawn. Organomercurial- and progesterone-induced maturations have many features in common: they do not occur when the inducer is injected into the oocytes, they require the presence of Ca++ in the medium, they are inhibited by cycloheximide but not by actinomycin D. In both cases, the maturation producing factor and the pseudomaturation inducing factor are produced. Organomercurial-treated oocytes react normally to activating stimuli; their protein synthesis increases, but uptake of amino acids is strongly inhibited. Progesterone and p-hydroxymercuriphenyl-sulfonate act synergically in inducing maturation. The main difference between the two agents is that p-hydroxymercuriphenylsulfonate must act for several hours, whereas, short contact with progesterone is sufficient to induce maturation.

Animals↗