Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collec...Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction （RT- PCR）, quantitative RT-PCR （QRT-PCR）, immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results： VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion： The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.展开更多
Objective To identify genes that involved in spermatogenesis. Methods In order to screen the testis-specific genes, testes cDNA samples from BALB/c mice of different postnatal days （days 4, 9, 18, 35, 54 and 6 months...Objective To identify genes that involved in spermatogenesis. Methods In order to screen the testis-specific genes, testes cDNA samples from BALB/c mice of different postnatal days （days 4, 9, 18, 35, 54 and 6 months） were performed with mouse whole genome Affymetrix chip. The characteristics of the selected gene were analyzed by various bioinformatic tools. The expression profile of the selected gene was identified by RT-PCR. Results By analyzing the hybridization signals, a gene with a differential expression in the developmental stages of testis was identified. This gene was designated as TSF22. The full length cDNA of 1 597 bp contained an open reading frame of 570 bp which encoded a putative protein of 190 amino acids and a molecular weight of 22.106 kD. RT-PCR analysis revealed that TSF22 mRNA was exclusively expressed in mice testis. Conclusions TSF22, functions as a testis-specific transcription factor, may play important roles during spermatogenesis.展开更多
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...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.展开更多
Objective To investigate the expression and the distribution of Dickkopf-likel （Dkkll） protein during the development of mouse testis. Methods Testes eDNA samples from BALB/c mice in different postnatal days were hy...Objective To investigate the expression and the distribution of Dickkopf-likel （Dkkll） protein during the development of mouse testis. Methods Testes eDNA samples from BALB/c mice in different postnatal days were hybridized with mouse whole genome affymetrix chip to screen the spermatogenesisrelated genes. The characteristics of the selected genes were analyzed by various bioinformatics tools. The mRNA expression of Dkkll at different stages of testis development and different tissues in mouse were analyzed by RT-PCR. The protein localization of Dkkll in mouse testis was assesed by immunohistoehemistry. Results By analyzing the gene chip signals of mouse testis aged 4 d, 9 d, 18 d, 35 d, 54 d and 6 months, Dkkll was identified with a differential expression in the develop- mental stages of testis. RT-PCR analysis showed that the expression of Dkkll mRNA was firstly detected on 15 d testis tissue and gradually upregulated during the testis developing to the adult stage. The Dkkll protein was predominantly located in spermatocytes and round spermatids in mouse testis. Conclusion The expression of Dkkll is gradually upregulated during the development of mouse testis and corresponds to the mouse spermatogenesis. It may play a critical role in male mammalian spermatogenesis.展开更多
基金We would like to thank Mr Jian-Rong Zhang, Mr Li-Bing Zhang and Dr Zhen-Dong Yu for technical assistance. This work was supported by grants from the National Natural Science Foundation of China （No. 30500543）, Ministry of Education ＂985 project＂ （No. 985-2-054-29）, and Shenzhen Foundation of Science ＆ Technology （JH200505270413B）.
文摘Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction （RT- PCR）, quantitative RT-PCR （QRT-PCR）, immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results： VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion： The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.
基金This study was supported by grants from Chinese Natural Science Funds(No.30471728,No.30500543)Natural Science Funds of Guangdong Province(No.04007303)
文摘Objective To identify genes that involved in spermatogenesis. Methods In order to screen the testis-specific genes, testes cDNA samples from BALB/c mice of different postnatal days （days 4, 9, 18, 35, 54 and 6 months） were performed with mouse whole genome Affymetrix chip. The characteristics of the selected gene were analyzed by various bioinformatic tools. The expression profile of the selected gene was identified by RT-PCR. Results By analyzing the hybridization signals, a gene with a differential expression in the developmental stages of testis was identified. This gene was designated as TSF22. The full length cDNA of 1 597 bp contained an open reading frame of 570 bp which encoded a putative protein of 190 amino acids and a molecular weight of 22.106 kD. RT-PCR analysis revealed that TSF22 mRNA was exclusively expressed in mice testis. Conclusions TSF22, functions as a testis-specific transcription factor, may play important roles during spermatogenesis.
基金This study was supported by grants from National Natural Science Foundation of China (No.30700824, No.30770810)973 Program(2008CB517412)Guangdong Natural Science Foundation (No.7008952)
文摘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.
基金supported by the Medical Research Foundation of Guangdong Province(B2014426)the Qingyuan Foundation of Science&Technology(2012B011204127)+1 种基金the National Natural Science Foundation of China(No.81170613,No.81270740)Shenzhen Basic Research Funds for Distinguished Young Scientists(JC201005260216A)
文摘Objective To investigate the expression and the distribution of Dickkopf-likel （Dkkll） protein during the development of mouse testis. Methods Testes eDNA samples from BALB/c mice in different postnatal days were hybridized with mouse whole genome affymetrix chip to screen the spermatogenesisrelated genes. The characteristics of the selected genes were analyzed by various bioinformatics tools. The mRNA expression of Dkkll at different stages of testis development and different tissues in mouse were analyzed by RT-PCR. The protein localization of Dkkll in mouse testis was assesed by immunohistoehemistry. Results By analyzing the gene chip signals of mouse testis aged 4 d, 9 d, 18 d, 35 d, 54 d and 6 months, Dkkll was identified with a differential expression in the develop- mental stages of testis. RT-PCR analysis showed that the expression of Dkkll mRNA was firstly detected on 15 d testis tissue and gradually upregulated during the testis developing to the adult stage. The Dkkll protein was predominantly located in spermatocytes and round spermatids in mouse testis. Conclusion The expression of Dkkll is gradually upregulated during the development of mouse testis and corresponds to the mouse spermatogenesis. It may play a critical role in male mammalian spermatogenesis.
