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首页> 外文期刊>Cell Research >Identification of differentially expressed genes of primary spermatoeyte against round spermatid isolated from human testis using the laser capture microdissection technique
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Identification of differentially expressed genes of primary spermatoeyte against round spermatid isolated from human testis using the laser capture microdissection technique

机译：利用激光捕获显微切割技术鉴定从人睾丸分离的针对圆形精子的原代精原细胞的差异表达基因

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摘要

The method of laser capture microdissection (LCM) combined with suppiessive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differentiation and development. In the present study, over 10,000 primary spermatocytes and round speimatid cells were successfully isolated by LCM Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed.The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1 7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals. Partial sequence of cDNAs prepared from each clone was determined with an overall success rate of 84,4%. Genes encoding cytochrome c oxidase II and the rescue factor-humanin were most frequently expressed in primary spermatocytes, suggesting their roles involved in meiosis.
机译：开发了激光捕获显微切割（LCM）结合超减减杂交（SSH）的方法，以从人睾丸切片中分离特定的生殖细胞，并鉴定在分化和发育过程中表达的基因。本研究成功地通过LCM成功分离了10,000多个原代精子细胞和圆形皮细胞，利用原代精子细胞和圆形精子的cDNAs构建了原代精子细胞SSH cDNA文库，随机从75个样本中分离出cDNA的平均插入大小。挑选的白色克隆为500 bp，范围从250 bp到1 7 kb。使用点印迹法，总共检查了421个克隆，从而鉴定出390个发射强信号的阳性克隆。确定从每个克隆制备的cDNA的部分序列，总成功率为84.4％。编码细胞色素C氧化酶II和救援因子-人类蛋白的基因最常在原代精母细胞中表达，表明它们参与减数分裂的作用。

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