Yu Zhendong, Wu Bo, Tang Aifa, Chen Jing, Guo Xin, Qin Jie, Gui Yaoting, Cai Zhiming
Department of Clinical Laboratory, Peking University Shenzhen Hospital, Shenzhen, 518036, China.
J Huazhong Univ Sci Technolog Med Sci. 2009 Oct;29(5):535-9. doi: 10.1007/s11596-009-0502-9. Epub 2009 Oct 11.
In order to identify novel genes involved in spermatogenesis, testis cDNA samples from Balb/C mice of different postnatal days were hybridized with the whole mouse genome Affymetrix chip to screen the testis-specific genes. The characteristics of the selected genes were analyzed by RT-PCR as well as other bioinformatic tools. A novel differentially expressed testis-specific gene (GenBank Accession No: NM_029042) in the developmental stages of testes was identified, and named TSCPA. Cellular mapping prediction of TSCPA indicated that its protein was probably expressed in nuclei, and one putative domain (aa 332-377) was anchoring domain of cAMP-dependent type II PK. The result of subcellular localization of GFP-TSCPA fusion protein in Cos-7 cells showed that TSCPA protein was expressed in nuclei. RT-PCR analysis revealed that TSCPA was expressed specifically in mouse and human testis. TSCPA gene was expressed weakly in 21-day-old mouse testis and the expression was increased gradually from 38th day to 6th month of mouse testes. No expression of hTSCPA was found in cryptorchidism and Sertoli-cell-only syndrome patients. It was concluded that the expression profile of TSCPA in human and mice indicated that TSCPA might play an important role in spermatogenesis.
为了鉴定参与精子发生的新基因,将来自不同出生后天数的Balb/C小鼠的睾丸cDNA样本与全小鼠基因组Affymetrix芯片杂交,以筛选睾丸特异性基因。通过RT-PCR以及其他生物信息学工具分析所选基因的特征。在睾丸发育阶段鉴定出一个新的差异表达睾丸特异性基因(GenBank登录号:NM_029042),并命名为TSCPA。TSCPA的细胞定位预测表明其蛋白可能在细胞核中表达,一个假定结构域(氨基酸332 - 377)是cAMP依赖性II型PK的锚定结构域。GFP - TSCPA融合蛋白在Cos - 7细胞中的亚细胞定位结果表明TSCPA蛋白在细胞核中表达。RT-PCR分析显示TSCPA在小鼠和人类睾丸中特异性表达。TSCPA基因在21日龄小鼠睾丸中表达较弱,从小鼠睾丸的第38天到第6个月表达逐渐增加。在隐睾症和唯支持细胞综合征患者中未发现hTSCPA的表达。得出结论,TSCPA在人和小鼠中的表达谱表明TSCPA可能在精子发生中起重要作用。