Possible germ cell-Sertoli cell interactions are critical for establishing appropriate expression levels for the Sertoli cell-specific MicroRNA, miR-202-5p, in human testis

Basic and Clinical Andrology
Ali A DabajaDarius A Paduch

Abstract

To examine human microRNA expression in fertile men and subsequently to compare expression patterns of miRNAs in fertile and infertile men, specifically men with Sertoli Cell Only (SCO) histopathology. Testicular tissues from men with azoospermia and SCO, as well as those of men with normal spermatogenesis, were analyzed. MicroRNA was isolated using the miRCURY™ RNA Purification Kit. A miRCURY LNA™ Universal RT system was used for detection of microRNA by quantitative real-time PCR. MicroRNA localization was performed by in situ hybridizations (ISH) on formalin-fixed paraffin embedded (FFPE) tissue utilizing miRCURY LNA™ microRNA ISH technology. Statistical analysis was performed by GenEx V5.0. MicroRNA expression was determined for 13 normal fertile men and 5 men with the confirmed diagnosis of diffuse SCO. MiR-202-5p expression was reduced by 17-fold (P < 0.00001) in tissue from SCO men compared to normal. MiR-34c-5p was reduced by 346-fold (P < 0.00001), miR-10b was reduced 18-fold (P < 0.00001), miR-191 was reduced 20-fold (P = 0.001) and miR-126 was reduced 40-fold (P < 0.00001)) in tissues from SCO compared to normal fertile men. Using ISH, miR-202-5p was localized to Sertoli cells of men with normal spermatogenesis, but ...Continue Reading

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Citations

Mar 5, 2016·Journal of Assisted Reproduction and Genetics·Sezgin GunesRamazan Asci
Jul 15, 2015·The Journal of Endocrinology·Ian S McLennan, Michael W Pankhurst
May 5, 2016·Seminars in Cell & Developmental Biology·Filipe Tenorio Lira NetoMarc Goldstein
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Aug 19, 2021·Journal of Gynecology Obstetrics and Human Reproduction·Neda SaebniaAhmad Reza Bahrami

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Methods Mentioned

BETA
column chromatography
PCR
light microscopy

Software Mentioned

GenEX
LightCycler 480
Roche

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