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小鼠细胞周期蛋白B2 cDNA的分离以及细胞周期蛋白B1和B2在卵子发生、精子发生和早期胚胎发生过程中表达的谱系和时间特异性表征。

Isolation of the murine cyclin B2 cDNA and characterization of the lineage and temporal specificity of expression of the B1 and B2 cyclins during oogenesis, spermatogenesis and early embryogenesis.

作者信息

Chapman D L, Wolgemuth D J

机构信息

Department of Genetics and Development, Columbia University College of Physicians and Surgeons, New York, NY 10032.

出版信息

Development. 1993 May;118(1):229-40. doi: 10.1242/dev.118.1.229.

Abstract

A cDNA encoding the murine cyclin B2 (cycB2) was isolated from an adult mouse testis cDNA library as part of studies designed to identify cyclins involved in murine germ cell development. This cycB2 cDNA was then used to examine the pattern of cycB2 expression during male and female germ cell development and in early embryogenesis, and to compare this expression with the previously characterized expression of cycB1. A single 1.7 kb cycB2 transcript was detected by northern blot hybridization analysis of total RNA isolated from midgestation embryos and various adult tissues. Northern blot and in situ hybridization analyses revealed that cycB2 expression in the testis was most abundant in the germ cells, specifically in pachytene spermatocytes. This is in contrast to the highest levels of expression of cycB1 being present in early spermatids. In situ analysis of the ovary revealed cycB2 transcripts in both germ cells and somatic cells, specifically in the oocytes and granulosa cells of growing and mature follicles. The pattern of cycB1 and cycB2 expression in ovulated and fertilized eggs was also examined. While the steady state level of cycB1 and cycB2 signal remained constant in oocytes and ovulated eggs, signal of both appeared to decrease following fertilization. In addition, both cycB1 and cycB2 transcripts were detected in the cells of the inner cell mass and the trophectoderm of the blastocyst. These results demonstrate that there are lineage- and developmental-specific differences in the pattern of the B cyclins in mammalian germ cells, in contrast to the co-expression of B cyclins in the early conceptus.

摘要

作为旨在鉴定参与小鼠生殖细胞发育的细胞周期蛋白的研究的一部分,从成年小鼠睾丸cDNA文库中分离出编码小鼠细胞周期蛋白B2(cycB2)的cDNA。然后使用该cycB2 cDNA来检测cycB2在雄性和雌性生殖细胞发育以及早期胚胎发生过程中的表达模式,并将该表达与先前表征的cycB1的表达进行比较。通过对从中期妊娠胚胎和各种成年组织中分离的总RNA进行Northern印迹杂交分析,检测到单一的1.7 kb cycB2转录本。Northern印迹和原位杂交分析显示,睾丸中cycB2的表达在生殖细胞中最为丰富,特别是在粗线期精母细胞中。这与早期精子细胞中cycB1的最高表达水平形成对比。卵巢的原位分析显示,生殖细胞和颗粒细胞中均有cycB2转录本,特别是在生长卵泡和成熟卵泡的卵母细胞和颗粒细胞中。还检查了排卵和受精卵中cycB1和cycB2的表达模式。虽然cycB1和cycB2信号的稳态水平在卵母细胞和排卵卵中保持恒定,但受精后两者的信号似乎都降低了。此外,在囊胚的内细胞团和滋养外胚层细胞中均检测到cycB1和cycB2转录本。这些结果表明,与早期概念中B细胞周期蛋白的共表达相反,哺乳动物生殖细胞中B细胞周期蛋白的模式存在谱系和发育特异性差异。

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