{"title":"不育男性睾丸特异性组蛋白2B的蛋白质组学和遗传学解剖揭示了其对减数分裂和精子活力的贡献","authors":"Aniket Patankar M.Sc. , Digumarthi V.S. Sudhakar Ph.D. , Rahul Gajbhiye M.B.B.S., Ph.D. , Suchitra Surve M.D. , Kumarasamy Thangaraj Ph.D. , Priyanka Parte Ph.D.","doi":"10.1016/j.xfss.2022.07.003","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>To investigate testis-specific histone 2B (TSH2B) and its gene anomalies in infertile men.</p></div><div><h3>Design</h3><p>Case-control study.</p></div><div><h3>Setting</h3><p>Basic science laboratory.</p></div><div><h3>Patient(s)</h3><p>Fertile and infertile men.</p></div><div><h3>Intervention(s)</h3><p>Not applicable.</p></div><div><h3>Main Outcome Measure(s)</h3><p><span><span>The histone and protamine<span> status of sperm was studied by aniline blue and chromomycin A3 staining, respectively. Testis-specific </span></span>histone 2B<span>, total H2B, and phosphorylated TSH2B (pTSH2B) were estimated by Western blot analysis<span>. The frequency of genetic polymorphisms and rare variants in </span></span></span><span><em>H2BC1</em></span><span> was studied by Sanger sequencing. Phosphosites on TSH2B in sperm were identified by reverse-phase high-performance liquid chromatography purification of TSH2B followed by mass spectrometric analysis.</span></p></div><div><h3>Result(s)</h3><p>Aniline blue and chromomycin A3 staining revealed significantly higher histone retention and low protamine in sperm of infertile men. Sperm TSH2B and total H2B levels were significantly lower in oligozoospermic and oligoasthenozoospermic men (in both groups). The TSH2B levels were comparable in asthenozoospermic men; however, the pTSH2B level was significantly low. The <em>H2BC1</em><span> gene sequencing identified 6 variants, of which 2 are rare variants (rs368672899 and rs544942090) and 4 (rs4711096, rs4712959, rs4712960 and rs4712961) are single nucleotide polymorphisms. Minor allele frequency of 5′-untranslated region variant rs4711096 was significantly lower in infertile men (OR = 0.65). The rare nonsynonymous variant, rs368672899, p.Ser5Pro was seen in 1 oligoasthenoteratozoospermic individual. Interestingly, mass spectrometric analysis identified a site on TSH2B to bear a phosphate group in the sperm of fertile men.</span></p></div><div><h3>Conclusion(s)</h3><p><span>Our study reveals a defect in the replacement of somatic histones with testis-specific variants in infertile men. Chromatin compaction positively correlates with sperm motility, which is suggestive of its utility in diagnostic </span>semen analysis of infertile individuals. Our observations with TSH2B and its cognate gene in sperm of infertile men indicate an essential role for TSH2B in meiosis and its phosphorylation in sperm motility, respectively.</p></div>","PeriodicalId":73012,"journal":{"name":"F&S science","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2022-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Proteomic and genetic dissection of testis-specific histone 2B in infertile men reveals its contribution to meiosis and sperm motility\",\"authors\":\"Aniket Patankar M.Sc. , Digumarthi V.S. Sudhakar Ph.D. , Rahul Gajbhiye M.B.B.S., Ph.D. , Suchitra Surve M.D. , Kumarasamy Thangaraj Ph.D. , Priyanka Parte Ph.D.\",\"doi\":\"10.1016/j.xfss.2022.07.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><p>To investigate testis-specific histone 2B (TSH2B) and its gene anomalies in infertile men.</p></div><div><h3>Design</h3><p>Case-control study.</p></div><div><h3>Setting</h3><p>Basic science laboratory.</p></div><div><h3>Patient(s)</h3><p>Fertile and infertile men.</p></div><div><h3>Intervention(s)</h3><p>Not applicable.</p></div><div><h3>Main Outcome Measure(s)</h3><p><span><span>The histone and protamine<span> status of sperm was studied by aniline blue and chromomycin A3 staining, respectively. Testis-specific </span></span>histone 2B<span>, total H2B, and phosphorylated TSH2B (pTSH2B) were estimated by Western blot analysis<span>. The frequency of genetic polymorphisms and rare variants in </span></span></span><span><em>H2BC1</em></span><span> was studied by Sanger sequencing. Phosphosites on TSH2B in sperm were identified by reverse-phase high-performance liquid chromatography purification of TSH2B followed by mass spectrometric analysis.</span></p></div><div><h3>Result(s)</h3><p>Aniline blue and chromomycin A3 staining revealed significantly higher histone retention and low protamine in sperm of infertile men. Sperm TSH2B and total H2B levels were significantly lower in oligozoospermic and oligoasthenozoospermic men (in both groups). The TSH2B levels were comparable in asthenozoospermic men; however, the pTSH2B level was significantly low. The <em>H2BC1</em><span> gene sequencing identified 6 variants, of which 2 are rare variants (rs368672899 and rs544942090) and 4 (rs4711096, rs4712959, rs4712960 and rs4712961) are single nucleotide polymorphisms. Minor allele frequency of 5′-untranslated region variant rs4711096 was significantly lower in infertile men (OR = 0.65). The rare nonsynonymous variant, rs368672899, p.Ser5Pro was seen in 1 oligoasthenoteratozoospermic individual. Interestingly, mass spectrometric analysis identified a site on TSH2B to bear a phosphate group in the sperm of fertile men.</span></p></div><div><h3>Conclusion(s)</h3><p><span>Our study reveals a defect in the replacement of somatic histones with testis-specific variants in infertile men. Chromatin compaction positively correlates with sperm motility, which is suggestive of its utility in diagnostic </span>semen analysis of infertile individuals. Our observations with TSH2B and its cognate gene in sperm of infertile men indicate an essential role for TSH2B in meiosis and its phosphorylation in sperm motility, respectively.</p></div>\",\"PeriodicalId\":73012,\"journal\":{\"name\":\"F&S science\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"F&S science\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2666335X22000477\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"F&S science","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666335X22000477","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Proteomic and genetic dissection of testis-specific histone 2B in infertile men reveals its contribution to meiosis and sperm motility
Objective
To investigate testis-specific histone 2B (TSH2B) and its gene anomalies in infertile men.
Design
Case-control study.
Setting
Basic science laboratory.
Patient(s)
Fertile and infertile men.
Intervention(s)
Not applicable.
Main Outcome Measure(s)
The histone and protamine status of sperm was studied by aniline blue and chromomycin A3 staining, respectively. Testis-specific histone 2B, total H2B, and phosphorylated TSH2B (pTSH2B) were estimated by Western blot analysis. The frequency of genetic polymorphisms and rare variants in H2BC1 was studied by Sanger sequencing. Phosphosites on TSH2B in sperm were identified by reverse-phase high-performance liquid chromatography purification of TSH2B followed by mass spectrometric analysis.
Result(s)
Aniline blue and chromomycin A3 staining revealed significantly higher histone retention and low protamine in sperm of infertile men. Sperm TSH2B and total H2B levels were significantly lower in oligozoospermic and oligoasthenozoospermic men (in both groups). The TSH2B levels were comparable in asthenozoospermic men; however, the pTSH2B level was significantly low. The H2BC1 gene sequencing identified 6 variants, of which 2 are rare variants (rs368672899 and rs544942090) and 4 (rs4711096, rs4712959, rs4712960 and rs4712961) are single nucleotide polymorphisms. Minor allele frequency of 5′-untranslated region variant rs4711096 was significantly lower in infertile men (OR = 0.65). The rare nonsynonymous variant, rs368672899, p.Ser5Pro was seen in 1 oligoasthenoteratozoospermic individual. Interestingly, mass spectrometric analysis identified a site on TSH2B to bear a phosphate group in the sperm of fertile men.
Conclusion(s)
Our study reveals a defect in the replacement of somatic histones with testis-specific variants in infertile men. Chromatin compaction positively correlates with sperm motility, which is suggestive of its utility in diagnostic semen analysis of infertile individuals. Our observations with TSH2B and its cognate gene in sperm of infertile men indicate an essential role for TSH2B in meiosis and its phosphorylation in sperm motility, respectively.