{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,25]],"date-time":"2026-10-25T05:49:59Z","timestamp":1792907399180,"version":"4.5.0"},"reference-count":44,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2018,2,22]],"date-time":"2018-02-22T00:00:00Z","timestamp":1519257600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000066","name":"National Institute of Environmental Health Sciences","doi-asserted-by":"publisher","award":["P42 ES013660"],"award-info":[{"award-number":["P42 ES013660"]}],"id":[{"id":"10.13039\/100000066","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Andrology"],"published-print":{"date-parts":[[2018,3]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Sperm <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> is a sensitive monitoring endpoint for male reproductive toxicants, and a potential biomarker to assess male infertility and sperm quality. However, isolation of sperm <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> is a challenging procedure due to the heterogeneous population of cells present in the ejaculate, the low yield of <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> per spermatozoon, and the absence of 18S and 28S ribosomal <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> subunits. The unique biology of spermatozoa has created some uncertainty in the field about <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> isolation methods, indicating the need for rigorous quality control checks to ensure reproducibility of data generated from sperm <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content>. Therefore, we developed a reliable and effective protocol for <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> isolation from rat and human spermatozoa that delivers highly purified and intact <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content>, verified using <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content>\u2010specific electrophoretic chips and molecular biology approaches such as <jats:styled-content style=\"fixed-case\">RT<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> and Western blot analysis. The sperm <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> isolation technique was optimized using rat spermatozoa and then adapted to human spermatozoa. Three steps in the sperm isolation procedure, epididymal fluid collection, sperm purification, and spermatozoon <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> extraction, were evaluated and assessed. The sperm <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> extraction methodology consists of collection of rat epididymal fluid with repeated needle punctures of the epididymis, somatic cell elimination using detergent\u2010based somatic cell lysis buffer (<jats:styled-content style=\"fixed-case\">SCLB<\/jats:styled-content>) and the use of <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> isolation Kit. Rat sperm heads are more resistant to disruption than human spermatozoa, necessitating the addition of mechanical lysis with microbeads and heat in the rat protocol, whereas the human sperm protocol only required lysis buffer. In conclusion, this methodology results in reliable and consistent isolation of high\u2010quality sperm <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content>. Using this technique will aid in translation of data collected from animal models, and reproducibility of clinical assessment of male factor fertility using <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> molecular biomarkers.<\/jats:p>","DOI":"10.1111\/andr.12471","type":"journal-article","created":{"date-parts":[[2018,2,23]],"date-time":"2018-02-23T06:08:49Z","timestamp":1519366129000},"page":"374-383","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":44,"title":["High\u2010quality human and rat spermatozoal <scp>RNA<\/scp> isolation for functional genomic studies"],"prefix":"10.1111","volume":"6","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8135-9730","authenticated-orcid":false,"given":"E.","family":"Bianchi","sequence":"first","affiliation":[{"name":"Division of Urology Rhode Island Hospital Providence RI USA"},{"name":"Department of Pathology and Laboratory Medicine Brown University Providence RI USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"Stermer","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Brown University Providence RI USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"Boekelheide","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Brown University Providence RI USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Sigman","sequence":"additional","affiliation":[{"name":"Division of Urology Rhode Island Hospital Providence RI USA"},{"name":"Department of Pathology and Laboratory Medicine Brown University Providence RI USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S. 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