Advances in single-cell technology have enabled the detailed mapping of testicular cell transcriptomes,which is essential for understanding spermatogenesis.However,the fragmented nature of age-specific data from vario...Advances in single-cell technology have enabled the detailed mapping of testicular cell transcriptomes,which is essential for understanding spermatogenesis.However,the fragmented nature of age-specific data from various literature sources has hindered comprehensive analysis.To overcome this,the Human Testis Database(HumanTestisDB)was developed,consolidating multiple human testicular sequencing datasets to address this limitation.Through extensive investigation,38 unique cell types were identified,providing a detailed perspective on cellular variety.Furthermore,the database systematically categorizes samples into eight developmental stages,offering a structured framework to comprehend the temporal dynamics of testicular development.Each stage features comprehensive maps of cell-cell interactions,elucidating the complex communication network inside the testicular microenvironment at particular developmental stages.Moreover,by facilitating comparisons of interactions among various cell types at different stages,the database enables examining alterations that occur during critical transitions in spermatogenesis.HumanTestisDB,available at https://shalab.njmu.edu.cn/humantestisdb,offers vital insights into testicular transcriptomes and cellular interactions,serving as an essential resource for advancing research in reproductive biology.展开更多
Aim: To investigate the roles of the BGR-like gene in testicular development/spermatogenesis. Methods: A human testis cDNA microarray was hybridized with probes from human adult testes and embryo testes. The different...Aim: To investigate the roles of the BGR-like gene in testicular development/spermatogenesis. Methods: A human testis cDNA microarray was hybridized with probes from human adult testes and embryo testes. The differentially expressed clones were sequenced and analyzed. Expression of the BGR-like gene was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Results: A new gene exhibiting 50-fold difference in expression level between adult and fetal human testes was cloned and named the BGR-like gene. The cDNA consisted of 2500 nucleotides and had an open reading frame of 1437 nucleotides encoding a putative protein of 497 amino acid residues. Homologous comparison showed that the BGR-like gene was a new alternative splicing variant of the BGR gene and had sequence homology with the bubblegum gene of human, mouse, rat and Drosophilia. Protein motif analysis of the BGR-like gene revealed that it contained a conserved adenosine monophosphate (AMP)-binding domain and a fatty acyl-CoA synthetase signature motif which existed in all acyl-CoA synthetases. The BGR-like gene transcript was imperceptibly expressed in human fetal testes, highly in human adult testes and moderately in elderly testes and human Leydig cells. RT-PCR-based tissue distribution experiments showed that the BGR-like gene was exclusively expressed in testes and was a testes-specific isoform of the BGR gene. A BGR-like gene transcript was not detected in some azoospermic testes. Conclusion: The BGR-like gene may play an important role in spermatogenesis/testicular development and may be correlated with male infertility.展开更多
Aim: To investigate the stage-specific localization of transforming growth factor (TGF) β1 and β3 during spermatogenesis in adult human testis. Methods: The localization of TGFβ1 and β3 was investigated by immunoh...Aim: To investigate the stage-specific localization of transforming growth factor (TGF) β1 and β3 during spermatogenesis in adult human testis. Methods: The localization of TGFβ1 and β3 was investigated by immunohis tochemical staining method employing specific polyclonal antibodies. Results: Both TGFβ1 and β3 and their recep tors were preponderant in the Leydig celis. TGFβ1 could not be detected in the seminiferous tubules. TGFβ3 and TGFβ-Receptor (R) I were mainly seen in the elongated spermatids, while TGFβ-RⅡ in the pachytene spermatocytes and weak in the spermatogonia, spermatids and Sertoli celis. Only TGFβ-RⅡ was detected in the Sertoli celis. TGFβ3, TGFβ-RⅠ and TGFβ-RⅡ showed a staining pattern dependent upon the stages of the seminiferous epithelium cycle. Conclusion: TGFβ isoforms and their receptors are present in the somatic and germ celis of the adult human testis, suggesting their involvement in the regulation of spermatogenesis.展开更多
Aim:To identify genes related to the human testis development by substrate hybridization technique.Methods:A human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal ...Aim:To identify genes related to the human testis development by substrate hybridization technique.Methods:A human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription reactions.The differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank database.Multi-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to deter- mine the tissue expression profile of cul-3b.Results:Cul-3b,a novel CUL-3 transcript variant,was identified.The expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ones.Cul-3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites (IRESes) in the 5'-UTR.These led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict circumstances.Additionally,cul-3b expressed ubiquitously in human tissues according to multi-tissue RT-PCR.Conclusion:Cul-3b is a novel transcript variant of CUL-3,which may be important not only for the development of human testis but also for that of other organs.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.82122025 to Yan Yuan and 82221005 to Jiahao Sha)the National Key R&D Program(Grant Nos.2022YFC2702800 to Yan Yuan,2021YFC2700302 to Jiahao Sha,and 2021YFC2700200 to Yiqiang Cui).
文摘Advances in single-cell technology have enabled the detailed mapping of testicular cell transcriptomes,which is essential for understanding spermatogenesis.However,the fragmented nature of age-specific data from various literature sources has hindered comprehensive analysis.To overcome this,the Human Testis Database(HumanTestisDB)was developed,consolidating multiple human testicular sequencing datasets to address this limitation.Through extensive investigation,38 unique cell types were identified,providing a detailed perspective on cellular variety.Furthermore,the database systematically categorizes samples into eight developmental stages,offering a structured framework to comprehend the temporal dynamics of testicular development.Each stage features comprehensive maps of cell-cell interactions,elucidating the complex communication network inside the testicular microenvironment at particular developmental stages.Moreover,by facilitating comparisons of interactions among various cell types at different stages,the database enables examining alterations that occur during critical transitions in spermatogenesis.HumanTestisDB,available at https://shalab.njmu.edu.cn/humantestisdb,offers vital insights into testicular transcriptomes and cellular interactions,serving as an essential resource for advancing research in reproductive biology.
文摘Aim: To investigate the roles of the BGR-like gene in testicular development/spermatogenesis. Methods: A human testis cDNA microarray was hybridized with probes from human adult testes and embryo testes. The differentially expressed clones were sequenced and analyzed. Expression of the BGR-like gene was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Results: A new gene exhibiting 50-fold difference in expression level between adult and fetal human testes was cloned and named the BGR-like gene. The cDNA consisted of 2500 nucleotides and had an open reading frame of 1437 nucleotides encoding a putative protein of 497 amino acid residues. Homologous comparison showed that the BGR-like gene was a new alternative splicing variant of the BGR gene and had sequence homology with the bubblegum gene of human, mouse, rat and Drosophilia. Protein motif analysis of the BGR-like gene revealed that it contained a conserved adenosine monophosphate (AMP)-binding domain and a fatty acyl-CoA synthetase signature motif which existed in all acyl-CoA synthetases. The BGR-like gene transcript was imperceptibly expressed in human fetal testes, highly in human adult testes and moderately in elderly testes and human Leydig cells. RT-PCR-based tissue distribution experiments showed that the BGR-like gene was exclusively expressed in testes and was a testes-specific isoform of the BGR gene. A BGR-like gene transcript was not detected in some azoospermic testes. Conclusion: The BGR-like gene may play an important role in spermatogenesis/testicular development and may be correlated with male infertility.
文摘Aim: To investigate the stage-specific localization of transforming growth factor (TGF) β1 and β3 during spermatogenesis in adult human testis. Methods: The localization of TGFβ1 and β3 was investigated by immunohis tochemical staining method employing specific polyclonal antibodies. Results: Both TGFβ1 and β3 and their recep tors were preponderant in the Leydig celis. TGFβ1 could not be detected in the seminiferous tubules. TGFβ3 and TGFβ-Receptor (R) I were mainly seen in the elongated spermatids, while TGFβ-RⅡ in the pachytene spermatocytes and weak in the spermatogonia, spermatids and Sertoli celis. Only TGFβ-RⅡ was detected in the Sertoli celis. TGFβ3, TGFβ-RⅠ and TGFβ-RⅡ showed a staining pattern dependent upon the stages of the seminiferous epithelium cycle. Conclusion: TGFβ isoforms and their receptors are present in the somatic and germ celis of the adult human testis, suggesting their involvement in the regulation of spermatogenesis.
文摘Aim:To identify genes related to the human testis development by substrate hybridization technique.Methods:A human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription reactions.The differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank database.Multi-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to deter- mine the tissue expression profile of cul-3b.Results:Cul-3b,a novel CUL-3 transcript variant,was identified.The expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ones.Cul-3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites (IRESes) in the 5'-UTR.These led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict circumstances.Additionally,cul-3b expressed ubiquitously in human tissues according to multi-tissue RT-PCR.Conclusion:Cul-3b is a novel transcript variant of CUL-3,which may be important not only for the development of human testis but also for that of other organs.
