Charbonneau M, Bonnec G, Boujard D
Laboratoire de Biologie et Génétique du Développement, URA 256 CNRS, Université de Rennes I, France.
Cell Differ Dev. 1990 Sep;31(3):197-206. doi: 10.1016/0922-3371(90)90132-g.
We examined the qualitative patterns of protein synthesis in fully grown prophase-blocked oocytes of Xenopus laevis and after meiosis reinitiation accompanying maturation of the oocytes. Newly synthesized proteins labelled with [35S]methionine were run on isoelectric focusing gels and further separated in the second dimension on SDS-polyacrylamide slab gels. Three types of maturation inducer were compared: progesterone, considered as the natural inducer of Xenopus oocyte maturation, hCG (human chorionic gonadotropin) and insulin. Three polypeptides with apparent molecular masses of 37 kDa (pI 4.7-4.8), 78 kDa (pI 4.7) and 138 kDa (pI 4.6-4.7) were found to be always synthesized in all three types of stimulation, while the synthesis of a fourth one (molecular mass 116 kDa, pI 4.7) was arrested during oocyte maturation. Moreover, when the follicular cells surrounding the oocytes were part of the stimulating pathway, which is the case during hCG-induced maturation, an additional polypeptide was synthesized by the oocytes (molecular mass 106 kDa, pI 6.0-6.2). This polypeptide was not synthesized during progesterone- or insulin-induced oocyte maturation, two types of stimulation which do not require the presence of the follicular cells. The biological significance of the hCG-induced polypeptide, not necessary for oocyte maturation, is discussed. On the other hand, the four other modifications in protein synthesis taking place during all three types of maturation-inducing stimulation appear to be necessary for oocyte maturation, since oocytes which failed to mature in response to stimulation always missed one or several of these four polypeptides.
我们研究了非洲爪蟾完全成熟的处于前期阻滞的卵母细胞以及伴随卵母细胞成熟的减数分裂重新启动后的蛋白质合成定性模式。用[35S]甲硫氨酸标记的新合成蛋白质在等电聚焦凝胶上进行电泳,然后在第二维的SDS-聚丙烯酰胺平板凝胶上进一步分离。比较了三种类型的成熟诱导剂:孕酮,被认为是非洲爪蟾卵母细胞成熟的天然诱导剂;人绒毛膜促性腺激素(hCG)和胰岛素。发现三种表观分子量分别为37 kDa(pI 4.7 - 4.8)、78 kDa(pI 4.7)和138 kDa(pI 4.6 - 4.7)的多肽在所有三种类型的刺激下均始终被合成,而第四种多肽(分子量116 kDa,pI 4.7)的合成在卵母细胞成熟过程中被阻断。此外,当围绕卵母细胞的卵泡细胞参与刺激途径时(在hCG诱导的成熟过程中就是这种情况),卵母细胞会合成一种额外的多肽(分子量106 kDa,pI 6.0 - 6.2)。在孕酮或胰岛素诱导的卵母细胞成熟过程中不合成这种多肽,这两种刺激类型不需要卵泡细胞的存在。讨论了hCG诱导的多肽对卵母细胞成熟并非必需的生物学意义。另一方面,在所有三种类型的成熟诱导刺激过程中发生的蛋白质合成的其他四种变化似乎对卵母细胞成熟是必需的,因为未能对刺激作出成熟反应的卵母细胞总是缺少这四种多肽中的一种或几种。