title: Phosphoramide mustard induces autophagy markers and mTOR inhibition prevents follicle loss due to phosphoramide mustard exposure.
abstract: Phosphoramide mustard (PM) is an ovotoxic metabolite of cyclophosphamide. Postnatal day 4 Fisher 344 rat ovaries were exposed to vehicle control (1% DMSO) or PM (60μM)±LY294002 or rapamycin for 2 or 4 d. Transmission election microscopy revealed abnormally large golgi apparatus and electron dense mitochondria in PM-exposed ovaries prior to and at the time of follicle depletion. PM exposure increased (P<0.05) mRNA abundance of Bbc3, Cdkn1a, Ctfr, Edn1, Gstp1, Nqo1, Tlr4, Tnfrsfla, Txnrd1 and decreased (P<0.05) Casp1 and Il1b after 4d. PM exposure increased (P<0.1) BECN1 and LAMP, decreased (P<0.1) ABCB1 and did not alter ABCC1 protein. LY294002 did not impact PM-induced ovotoxicity, but decreased ABCC1 and ABCB1 protein. Rapamycin prevented PM-induced follicle loss. These data suggest that the mammalian target of rapamycin, mTOR, may be a gatekeeper of PM-induced follicle loss.
authors.affiliation: Department of Animal Science, Iowa State University, Ames, IA, 50011, United States.
authors.forename: Jill A
authors.initials: JA
authors.lastname: Madden
authors.affiliation: Department of Animal Science, Iowa State University, Ames, IA, 50011, United States.
authors.forename: Porsha Q
authors.initials: PQ
authors.lastname: Thomas
authors.affiliation: Department of Animal Science, Iowa State University, Ames, IA, 50011, United States. Electronic address: akeating@iastate.edu.
authors.forename: Aileen F
authors.initials: AF
authors.lastname: Keating
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D018906:Antineoplastic Agents, Alkylating / Q000633:toxicity*; D001343:Autophagy / Q000187:drug effects*; D005260:Female; D006056:Golgi Apparatus / Q000187:drug effects / Q000648:ultrastructure; D066298:In Vitro Techniques; D008928:Mitochondria / Q000187:drug effects / Q000648:ultrastructure; D006080:Ovarian Follicle / Q000187:drug effects / Q000648:ultrastructure; D010053:Ovary / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D010752:Phosphoramide Mustards / Q000633:toxicity*; D011916:Rats, Inbred F344; D020123:Sirolimus / Q000494:pharmacology; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors*
chemical_list: D018906:Antineoplastic Agents, Alkylating; D010752:Phosphoramide Mustards; C030090:phosphoramide mustard; C546845:mTOR protein, rat; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus
keywords: Mammalian target of rapamycin; Ovotoxicity; Phosphatidylinositol-3 kinase; Phosphoramide mustard