High-quality human and rat spermatozoal RNA isolation for functional genomic studies

Andrology
E BianchiK Hwang

Abstract

Sperm RNA is a sensitive monitoring endpoint for male reproductive toxicants, and a potential biomarker to assess male infertility and sperm quality. However, isolation of sperm RNA is a challenging procedure due to the heterogeneous population of cells present in the ejaculate, the low yield of RNA per spermatozoon, and the absence of 18S and 28S ribosomal RNA subunits. The unique biology of spermatozoa has created some uncertainty in the field about RNA isolation methods, indicating the need for rigorous quality control checks to ensure reproducibility of data generated from sperm RNA. Therefore, we developed a reliable and effective protocol for RNA isolation from rat and human spermatozoa that delivers highly purified and intact RNA, verified using RNA-specific electrophoretic chips and molecular biology approaches such as RT-PCR and Western blot analysis. The sperm RNA 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 RNA extraction, were evaluated and assessed. The sperm RNA extraction methodology consists of collection of rat epididymal fluid with repeated needle pun...Continue Reading

References

Jan 16, 1989·Biochemical and Biophysical Research Communications·C A PessotL O Burzio
Apr 1, 1987·Human Reproduction·M WiklandP O Janson
Feb 15, 2001·Nature Cell Biology·P M WassarmanE S Litscher
Apr 17, 2002·The Journal of Urology·Jonathan P JarowUNKNOWN Male Infertility Best Practice Policy Committee of the American Urological Association Inc
Sep 21, 2002·Lancet·G Charles OstermeierStephen A Krawetz
Feb 27, 2004·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Hui WangJiahao Sha
May 14, 2004·Nature·G Charles OstermeierStephen A Krawetz
Sep 24, 2004·International Review of Cytology·Jean-Pierre DadouneMarie-Françoise Alfonsi
Sep 2, 2005·Nature Reviews. Genetics·Stephen A Krawetz
Feb 2, 2006·BMC Molecular Biology·Andreas SchroederThomas Ragg
Jul 7, 2007·Archives of Andrology·Robert GoodrichStephen A Krawetz
Aug 9, 2007·Nature Clinical Practice. Urology·Larry I Lipshultz, Dolores J Lamb
May 2, 2009·Disease Models & Mechanisms·Philip M Iannaccone, Howard J Jacob
May 26, 2009·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Claudia LalancetteStephen A Krawetz
Sep 18, 2009·Reproduction : the Official Journal of the Society for the Study of Fertility·David MillerDavid Iles
Jul 10, 2010·Theriogenology·Pranab J DasTerje Raudsepp
Jul 20, 2010·Reproductive Biomedicine Online·J M FeugangE Memili
Dec 17, 2010·Der Urologe. Ausg. A·F GottardoUNKNOWN World Health Organization
Oct 13, 2011·Human Reproduction·Stephen A KrawetzMichael P Diamond
Sep 21, 2012·Methods in Molecular Biology·Robert J GoodrichStephen A Krawetz
Jul 17, 2013·Human Reproduction Update·Meritxell JodarUNKNOWN Reproductive Medicine Network
Jul 23, 2013·Systems Biology in Reproductive Medicine·Shihong MaoStephen A Krawetz
Aug 22, 2013·Expert Opinion on Drug Metabolism & Toxicology·Sarah CampionFederico Goodsaid
Jul 22, 2014·Animal Reproduction Science·C M ShafeequeP A Azeez
Aug 1, 2014·Systems Biology in Reproductive Medicine·Shihong MaoStephen A Krawetz
Aug 5, 2014·The Journal of Urology·Andrew P GeorgiadisAlexander N Yatsenko
May 14, 2016·Genetics and Molecular Research : GMR·C M HanQ H Gao
Jul 29, 2016·Toxicological Sciences : an Official Journal of the Society of Toxicology·Edward DereKim Boekelheide
Sep 16, 2016·Biology of Reproduction·Andrew SchusterWei Yan

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Citations

Jul 7, 2018·Epigenomics·Brandon L PearsonDan Ehninger
Oct 1, 2019·Molecular Reproduction and Development·Malsawmhriatzuala JeremyVikas Kumar Roy
Feb 16, 2019·Toxicological Sciences : an Official Journal of the Society of Toxicology·Angela R StermerKim Boekelheide
May 11, 2021·Frontiers in Cell and Developmental Biology·Martin Roszkowski, Isabelle M Mansuy
Jun 27, 2021·Reproductive Toxicology·Enrica BianchiKathleen Hwang
Aug 28, 2021·Antioxidants·Zofia E MadejaNatalia Rozwadowska
Oct 9, 2021·Human Reproduction Update·Joana SantiagoMargarida Fardilha
Sep 30, 2021·Reproduction, Fertility, and Development·Brahim Halil GüngörGaffari Türk

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