机构地区:[1]Department of Obstetrics and Gynaecology, Prince of Wales Hospital The Chinese University of Hong Kong, Hong Kong, China [2]Department of Animal Sciences, Laiyang Agricultural College, Qingdao, Shandong 266109, China [3]Center for Human Reproduction, North Shore University Hospital NYU School of Medicine, Manhasset, NY 11030, USA [4]Li Ka Shing Institute of Health Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China
出 处:《Asian Journal of Andrology》2008年第2期219-226,共8页亚洲男性学杂志(英文版)
摘 要:Aim: To determine the possible roles of the t-complex testis expressed gene 5 (Tctex5) on sperm functions, the fulllength sequence of mRNA was studied and compared in the testis between the normal wild-type and the sterile t-haplotype mutant mice. Methods: We applied rapid amplification of cDNA ends, Northern blot and reverse transcription polymerase chain reaction to analyze the full length of Tctex5 mRNAs isolated from testes of the wild-type and the t-haplotype mice. Reverse transcription polymerase chain reaction was used to semi-quantitatively compare expression of Tctex5 transcripts in the 16 tissues and 9.5 day stage embryos in the wild-type mice. E-translation was applied to estimate the amino acid sequences. Results: One long and one short transcript of Tctex5 mRNA were discovered in mouse testis of wild-type (Tctex5^long-+ and Tctex5^short-+) and t-haplotype (Tctex5^long-+ and Tctex5^short-+) mice, respectively. Being enhanced only in the testis, Tctex5^long-+ had 17 point mutations and one 15-bp-deletion in the exon 1 region, comparing with the Tctex5^long-+, whereas the Tctex5^short-+ was similar to the Tctex5^short-+. The short isoforms of Tctex5 mRNAs in the two models encoded exactly the same peptides, but the long isoforms did not. The estimated peptide encoded by Tctex5^long-+ had significant mutations on putative sites of phosphorylation and PP1 binding. Conclusion: We established that mutations that occur in the Tctex5 long transcript of the t-haplotype mice are important for normal sperm function, whereas the short transcript of Tctex5 might have a conserved function among different tissues. (Asian J Androl 2008 Mar; 10: 219-226)Aim: To determine the possible roles of the t-complex testis expressed gene 5 (Tctex5) on sperm functions, the fulllength sequence of mRNA was studied and compared in the testis between the normal wild-type and the sterile t-haplotype mutant mice. Methods: We applied rapid amplification of cDNA ends, Northern blot and reverse transcription polymerase chain reaction to analyze the full length of Tctex5 mRNAs isolated from testes of the wild-type and the t-haplotype mice. Reverse transcription polymerase chain reaction was used to semi-quantitatively compare expression of Tctex5 transcripts in the 16 tissues and 9.5 day stage embryos in the wild-type mice. E-translation was applied to estimate the amino acid sequences. Results: One long and one short transcript of Tctex5 mRNA were discovered in mouse testis of wild-type (Tctex5^long-+ and Tctex5^short-+) and t-haplotype (Tctex5^long-+ and Tctex5^short-+) mice, respectively. Being enhanced only in the testis, Tctex5^long-+ had 17 point mutations and one 15-bp-deletion in the exon 1 region, comparing with the Tctex5^long-+, whereas the Tctex5^short-+ was similar to the Tctex5^short-+. The short isoforms of Tctex5 mRNAs in the two models encoded exactly the same peptides, but the long isoforms did not. The estimated peptide encoded by Tctex5^long-+ had significant mutations on putative sites of phosphorylation and PP1 binding. Conclusion: We established that mutations that occur in the Tctex5 long transcript of the t-haplotype mice are important for normal sperm function, whereas the short transcript of Tctex5 might have a conserved function among different tissues. (Asian J Androl 2008 Mar; 10: 219-226)
关 键 词:t-complex testis expressed gene 5 TRANSCRIPTS TESTIS mice
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