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PLoS One. 2019 May 23;14(5):e0216584. doi: 10.1371/journal.pone.0216584. eCollection 2019.

Spermatozoal large RNA content is associated with semen characteristics, sociodemographic and lifestyle factors.

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Division of Urology, Rhode Island Hospital, Providence, Rhode Island, United States of America.
Department of Pathology and Laboratory Medicine, Brown University, Providence, Rhode Island, United States of America.
Department of Epidemiology, School of Public Health, Brown University, Providence, Rhode Island, United States of America.
Department of Urology, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania, United States of America.


Semen analysis is one of the standard diagnostic tools currently used to assess male infertility and reproductive toxicity. However, semen analysis has a limited ability to separate fertile from infertile populations. Additional methods to detect impaired fertility are needed. The purpose of the present study was to evaluate how spermatozoal RNA content varies with sociodemographic and behavior/lifestyle factors, and to determine if spermatozoal large and small RNAs discriminate normal from abnormal spermatozoa. Semen specimens were collected from 133 men aged between 18 to 55 years undergoing semen analysis as part of couple infertility evaluation while 10 proven fertile donors were recruited as control group. Semen samples were classified as normal or abnormal according to World Health Organization (WHO) 2010 criteria. Sperm RNAs were extracted after somatic cells were lysed, and the association of large or small RNA content with semen quality and sociodemographic and behavioral/lifestyle factors was evaluated using a generalized additive model and one-way ANOVA. Inverse relationship was observed between large RNA content and sperm parameters such as sperm count, density and motility. Large RNA content per sperm was significantly increased in semen samples showing abnormal number of round cells. Furthermore, sperm motility was inversely associated with spermatozoal small RNA contents. Grouping donors by the number of semen abnormalities, we observed significant increased spermatozoal large and small RNA content in men with more than two semen abnormalities. Alcohol consumption was strongly associated with increased large RNA per sperm concentration after adjustment for age and BMI. Our study demonstrates a strong relationship between spermatozoal large RNA content and poor semen characteristics that may lead to a role in the assessment of male fertility, and may be used as an endpoint for reproductive toxicology risk assessment.

Conflict of interest statement

Kim Boekelheide is an occasional expert consultant for chemical and pharmaceutical companies; within the past 12 months he has consulted with TbAlliance, a non-profit pharmaceutical company developing anti-tuberculosis therapies, and has established an in-kind collaboration with Corteva Agrisciences. These activities are unrelated to the research and data reported in this publication, and are reported in the interest of full disclosure. Kim Boekelheide and Susan J. Hall both own stock in Semma Therapeutics (formerly CytoSolv, Inc.) a not publicly-traded small biotechnology start-up company involved in the development of a cell-based therapy for diabetes. This does not alter our adherence to PLOS ONE policies on sharing data and materials. The authors have no additional financial interests. Each author has read and approved of this manuscript submission and accepts responsibility for its content.

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