Inhibition of PTEN activates bovine non-growing follicles in vitro but increases DNA damage and reduces DNA repair response

抑制 PTEN 可在体外激活牛非生长卵泡,但会增加 DNA 损伤并降低 DNA 修复反应

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作者:Mila Maidarti, Yvonne L Clarkson, Marie McLaughlin, Richard A Anderson, Evelyn E Telfer

Methods

Bovine ovaries were obtained from abattoir-slaughtered heifers. Tissue fragments were exposed to either control medium alone or medium containing either 1 μM or 10 μM bpv(HOpic) for 24 h. Tissue fragments collected after 24 h were subjected to Akt quantification by Western blotting (six to nine fragments per group per experiment). Follicle stage and morphology were classified in remaining fragments. Immunohistochemical analysis included nuclear exclusion of FOXO3 as a marker of follicle activation, γH2AX as a marker of DNA damage, meiotic recombination 11 (MRE11), ataxia telangiectasia mutated (ATM), Rad51, breast cancer susceptibility 1 (BRCA1) and breast cancer susceptibility 2 (BRCA2) as DNA repair factors. A total of 29 550 follicles from three independent experiments were analysed. Main

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