title: Evaluation of boar sperm as a model system to study the mechanism of spermicidal activity of vanadocenes.
abstract: bis-cyclopentadienyl [Cp] complexes of vanadium(IV) or vanadocenes are rapid and potent inhibitors of human sperm motility with potential as a new class of contraceptive agents. We investigated the utility of boar sperm as a model system to study the mechanisms of drug action because boar sperm lacks phosphocreatine and creatine kinase activity, the essential components of the "phosphagen shuttle" system for human sperm motility. Two representative vanadocenes, vanadocene dichloride [VDC] and bis[pentamethylcyclopentadienyl] vanadium dichloride [VPMDC], in which the bis-Cp rings were substituted with five electron-donating methyl groups were evaluated. The concentration-dependent effects of VDC and VPMDC on spermicidal activity, axonemal dynein adenosine triphosphatase (ATPase) activity, and tyrosine phosphorylation of global sperm proteins were assessed by computer-assisted sperm analysis, spectrophotometry, and immunoblotting, respectively. Both the unsubstituted and the pentamethyl-substituted vanadocene induced rapid sperm immobilization (T(1/2) < 15 s). Substitution of the bis-Cp rings by five methyl groups augmented the SIA of VDC threefold. The EC(50) values for VDC and VPMDC were 2.1 and 0.76 microM, respectively. Spermicidal activity of vanadocenes was not associated with the inhibition of dynein ATPase(s) or increase in tyrosine phosphorylation of sperm proteins. These results suggest that the potent spermicidal activity of vanadocenes against boar sperm is mediated by a unique mechanism that is independent of dynein ATPase activity, phosphatase activity, and phosphocreatine/creatine kinase system. Therefore, boar sperm is a suitable model for further investigating the molecular mechanism of spermicidal action of vanadocenes.
authors.affiliation: Drug Discovery Program, Department of Reproductive Biology, Department of Biochemistry, Parker Hughes Institute, 2665 Long Lake Road, St. Paul, Minnesota 55113, USA. odcruz@ih.org
authors.forename: O J
authors.initials: OJ
authors.lastname: D'Cruz
authors.forename: A
authors.initials: A
authors.lastname: Vassilev
authors.forename: F M
authors.initials: FM
authors.lastname: Uckun
mesh_terms: D000818:Animals; D004305:Dose-Response Relationship, Drug; D004398:Dyneins / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D009942:Organometallic Compounds / Q000633:toxicity*; D013081:Sperm Motility / Q000187:drug effects*; D013089:Spermatocidal Agents / Q000633:toxicity*; D013094:Spermatozoa / Q000187:drug effects* / Q000201:enzymology; D013552:Swine; D017968:Vanadium Compounds / Q000633:toxicity
chemical_list: D009942:Organometallic Compounds; D013089:Spermatocidal Agents; D017968:Vanadium Compounds; C114984:bis(cyclopentadienyl)vanadium(IV); C030754:vanadocene dichloride; D004398:Dyneins