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 testisspecific 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 Acces-sion 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 subcel-lular 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-cellonly 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.
Objective To analyze the expression of SPACA4 in human and mice. Methods Testes cRNA samples from Balb/c mice of different postnatal days were performed with mouse affymetrix chip to screen the expression of SPACA4 in mice. Sub-quantitative RT-PCR and bioinformatic tools were used here to describe the expression profile of SPACA4 in mice and human. Results The results of gene chip analysis indicated that the expression of mSPACA4 began after d 35 of postnatal testis in mice. Sub-quantitative RT-PCR assay showed that SPACA4 gene expressed exclusively in mouse and human testis, and mouse mSPACA4 gene expressed after d 35 of postnatal testis that was consistency with the results of gene chip analysis. By bioinformatics analysis, mSPACA4 is located in cell membrane (34.8%) or plasma membrane (34.8%), the signal peptide cleavage site between position 19 and 20 amino acids, transmembrane region between 2-20 and 101-126 amino acids, respectively, on mSPACA4 protein. Conclusion mSPACA4 and hSPACA4 were testis-specific genes, and the expression of mSPACA4 begins after d 35 of postnatal testis in mice. SPACA4 is a candidate for targeting in a sperm-based contraceptive vaccine.
Ai-fa TANGZhen-dong YUYao-ting GUIXin GUOXian-xin LIWei-xiang LIUHui ZHUZhi-ming CAI
