title: Regulation of microcystin-LR-induced toxicity in mouse spermatogonia by miR-96.
abstract: Microcystin (MC)-LR is a cyclic heptapeptide that acts as a potent reproductive system toxin, especially by decreasing sperm quality through affecting spermatogonia. However, the molecular mechanisms of MC-induced spermatogonial cytotoxicity still remain unclear. The present study was designed to investigate changes in microRNA (miRNA) profiles and their potential functions in spermatogonia (GC-1 cell line) following treatment with MC-LR. With microarray analysis, 101 miRNAs were identified to be significantly altered in GC-1 cells treated with MC-LR. Among the 25 miRNAs associated with spermatogenesis, miR-96 was down-regulated most dramatically and thus selected for further functional analysis. Deleted-in azoospermia-associated protein 2 (DAZAP2) was predicted to have a binding sequence for miR-96 within its 3'-untranslated region. Fluorescent reporter assay confirmed that DAZAP2 was the target gene of miR-96. The expression of DAZAP2 decreased significantly when miR-96 was up-regulated. Consistently, down-regulation of miR-96 significantly increased the level of DAZAP2. Up-regulation of miR-96 promoted cell viability in GC-1 cells as a result of exposure to MC-LR. Our study suggested a crucial role for miR-96 in the regulation of cytotoxic effects of MC-LR in spermatogonia, which provides new perspectives in the diagnosis and treatment strategies for MC-induced male infertility.
authors.affiliation: Immunology and Reproduction Biology Laboratory, Medical School, Nanjing University , Nanjing 210093, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Zou
authors.initials: Z
authors.lastname: Xiang
authors.forename: Dongmei
authors.initials: D
authors.lastname: Li
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Han
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D008297:Male; D008387:Marine Toxins; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D052998:Microcystins / Q000633:toxicity*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D013093:Spermatogonia / Q000187:drug effects* / Q000378:metabolism
chemical_list: D008387:Marine Toxins; D035683:MicroRNAs; D052998:Microcystins; C541554:Mirn96 microRNA, mouse; D016601:RNA-Binding Proteins; C057862:cyanoginosin LR