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小鼠卵母细胞生长、成熟及植入前胚胎发育过程中的G蛋白基因表达。

G protein gene expression during mouse oocyte growth and maturation, and preimplantation embryo development.

作者信息

Williams C J, Schultz R M, Kopf G S

机构信息

Department of Obstetrics and Gynecology, University of Pennsylvania, Philadelphia 19104-6080, USA.

出版信息

Mol Reprod Dev. 1996 Jul;44(3):315-23. doi: 10.1002/(SICI)1098-2795(199607)44:3<315::AID-MRD5>3.0.CO;2-P.

Abstract

Fertilization in mammals initiates "egg activation," a series of events leading to embryo development. The signal transduction events that occur as a result of sperm-egg interactions and that initiate egg activation may be analogous to a ligand-receptor-effector pathway, but the details of this signaling pathway are poorly understood. Several lines of evidence support a role for guanine nucleotide-binding regulatory proteins (G proteins) in mammalian egg activation. Prior to initiating studies to examine further the role of specific G proteins in sperm-induced mouse egg activation, we needed to define the complement of G proteins expressed in the egg. Using a reverse transcription-polymerase chain reaction (RT-PCR) assay, the relative levels of mRNAs encoding specific G protein alpha, beta, and gamma subunits were determined in meiotically incompetent oocytes, fully-grown competent oocytes, metaphase II-arrested eggs, one-, two-, and eight-cell embryos, and blastocysts. mRNA transcripts representing all of the heterotrimeric G protein families were present at all of the stages examined, and all underwent significant changes in their patterns of expression. The following heterotrimeric G protein mRNA transcripts were present in oocytes, eggs, or preimplantation embryos: G alpha q family (q, 11, and 14), G alpha 12 family (12 and 13), G alpha i family (i1, i2, i3, t2, z, and s), beta subunits 1, 2, 4, and 5, and gamma subunits 2, 3, 5, and 7. A recently described large molecular weight G protein, G alpha h (Nakaoka et al., 1994: Science 264:1593-1596), was also present, G alpha 15, G alpha t1, G alpha olf, G alpha oA, G beta 3, G gamma 1, and G gamma 8 mRNA transcripts were not detected using this method. The most common pattern of expression observed was a maturation-associated decrease followed by an increase after the two-cell stage. Some transcripts, however, were expressed at low levels until the eight-cell to blastocyst stages, whereas others were expressed at high levels in the oocyte but following maturation declined and remained at a low level throughout preimplantation development.

摘要

哺乳动物的受精过程启动了“卵子激活”,这是一系列导致胚胎发育的事件。精子与卵子相互作用引发的信号转导事件以及启动卵子激活的过程,可能类似于配体-受体-效应器途径,但这条信号通路的具体细节尚不清楚。有几条证据支持鸟嘌呤核苷酸结合调节蛋白(G蛋白)在哺乳动物卵子激活中发挥作用。在开始进一步研究特定G蛋白在精子诱导的小鼠卵子激活中的作用之前,我们需要确定卵子中表达的G蛋白种类。使用逆转录-聚合酶链反应(RT-PCR)分析,测定了减数分裂无能的卵母细胞、完全成熟的有能卵母细胞、中期II期阻滞的卵子、一细胞、二细胞和八细胞胚胎以及囊胚中编码特定G蛋白α、β和γ亚基的mRNA的相对水平。在所检测的所有阶段都存在代表所有异三聚体G蛋白家族的mRNA转录本,并且它们的表达模式都发生了显著变化。以下异三聚体G蛋白mRNA转录本存在于卵母细胞、卵子或植入前胚胎中:Gαq家族(q、11和14)、Gα12家族(12和13)、Gαi家族(i1、i2、i3、t2、z和s)、β亚基1、2、4和5以及γ亚基2、3、5和7。最近描述的一种大分子G蛋白Gαh(中冈等人,1994年:《科学》264:1593 - 1596)也存在,未使用此方法检测到Gα15、Gαt1、Gαolf、GαoA、Gβ3、Gγ1和Gγ8的mRNA转录本。观察到的最常见表达模式是与成熟相关的下降,随后在二细胞阶段后增加。然而,一些转录本在八细胞到囊胚阶段之前表达水平较低,而另一些在卵母细胞中高水平表达,但在成熟后下降并在整个植入前发育过程中保持在低水平。

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