{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T07:58:14Z","timestamp":1791359894067,"version":"4.3.1"},"reference-count":46,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2019,3,28]],"date-time":"2019-03-28T00:00:00Z","timestamp":1553731200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2011\/19551\u20101"],"award-info":[{"award-number":["2011\/19551\u20101"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2011 \/ 19551\u20101"],"award-info":[{"award-number":["2011 \/ 19551\u20101"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["001"],"award-info":[{"award-number":["001"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Andrology"],"published-print":{"date-parts":[[2019,5]]},"abstract":"Abstract<\/jats:title>Background<\/jats:title>Sperm DNA<\/jats:styled-content> integrity is crucial for transmission of genetic information to future generations and DNA<\/jats:styled-content> damage can occur during chromatin packaging. Chromatin packaging involves the replacement of somatic nucleosomal histones by nuclear proteins called protamines. Protamine 1 (PRM<\/jats:styled-content>1) is transcribed and translated in spermatids of all mammals; however, protamine 2 (PRM<\/jats:styled-content>2) is transcribed in low levels in spermatids and it is not yet described in bull mature spermatozoa.<\/jats:p><\/jats:sec>Objectives<\/jats:title>The aim of this study was to assess gene and protein expression of PRM<\/jats:styled-content>2 and corroborate gene and protein expression of PRM<\/jats:styled-content>1 in bull spermatozoa and testis.<\/jats:p><\/jats:sec>Materials and methods<\/jats:title>For this purpose, absolute q\u2010RT<\/jats:styled-content>\u2010PCR<\/jats:styled-content> was performed to calculate the number of copies of PRM<\/jats:styled-content>1 and PRM<\/jats:styled-content>2 mRNA<\/jats:styled-content>s in bovine epididymal spermatozoa and testicular tissue. Western blot and mass spectrometry were performed to identify PRM<\/jats:styled-content>1 and PRM<\/jats:styled-content>2 in samples of bovine epididymal spermatozoa. Samples of bovine testicular tissue were collected to identify PRM<\/jats:styled-content>1 and PRM<\/jats:styled-content>2 by immunohistochemistry.<\/jats:p><\/jats:sec>Results<\/jats:title>We evaluated that the number of PRM<\/jats:styled-content>1 mRNA<\/jats:styled-content> copies was about hundred times higher than PRM<\/jats:styled-content>2 mRNA<\/jats:styled-content> copies in sperm and testicular samples (p\u00a0<\/jats:italic><\u00a0<\/jats:italic>0.0001). In addition, we estimated the PRM<\/jats:styled-content>1: PRM<\/jats:styled-content>2 ratio based on mRNA<\/jats:styled-content> number of copies. In spermatozoa, the ratio was 1: 0.014, and in testicle, the ratio was 1: 0.009. We also evaluated the immunolocalization for PRM<\/jats:styled-content>1 and PRM<\/jats:styled-content>2 in bovine testis, and both proteins were detected in spermatids. Western blot and mass spectrometry in bovine epididymal spermatozoa confirmed these results.<\/jats:p><\/jats:sec>Conclusion<\/jats:title>Our work identifies, for the first time, PRM<\/jats:styled-content>2 in bovine epididymal spermatozoa and in testis. Further studies are still needed to understand the role of PRM<\/jats:styled-content>2 on the chromatin of the spermatozoa and to verify how possible changes in PRM<\/jats:styled-content>2 levels may influence the bull fertility.<\/jats:p><\/jats:sec>","DOI":"10.1111\/andr.12610","type":"journal-article","created":{"date-parts":[[2019,3,28]],"date-time":"2019-03-28T14:38:04Z","timestamp":1553783884000},"page":"373-381","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Detection of protamine 2 in bovine spermatozoa and testicles"],"prefix":"10.1111","volume":"7","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8536-3123","authenticated-orcid":false,"given":"T. R. S.","family":"Hamilton","sequence":"first","affiliation":[{"name":"Department of Animal Reproduction School of Veterinary Medicine University of S\u00e3o Paulo Avenida Professor Doutor Orlando Marques de Paiva Sao Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"Sim\u00f5es","sequence":"additional","affiliation":[{"name":"Center for Natural and Human Sciences Federal University of ABC Santo Andr\u00e9 Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. M.","family":"Mendes","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction School of Veterinary Medicine University of S\u00e3o Paulo Avenida Professor Doutor Orlando Marques de Paiva Sao Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. D.","family":"Goissis","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction School of Veterinary Medicine University of S\u00e3o Paulo Avenida Professor Doutor Orlando Marques de Paiva Sao Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"Nakajima","sequence":"additional","affiliation":[{"name":"Process Development Laboratory Inovation Division Butantan Institute S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E. A. L.","family":"Martins","sequence":"additional","affiliation":[{"name":"Process Development Laboratory Inovation Division Butantan Institute S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. 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