title: Multivariate analysis of male reproductive function in Inpp5b-/- mice reveals heterogeneity in defects in fertility, sperm-egg membrane interaction and proteolytic cleavage of sperm ADAMs.
abstract: Past work indicated that sperm from mice deficient in the inositol polyphosphate 5-phosphatase Inpp5b have reduced ability to fertilize eggs in vitro and reduced epididymal proteolytic processing of the sperm protein A Disintegrin and A Metalloprotease 2 (ADAM2). On the basis of these data, our central working hypothesis was that reduced ADAM cleavage would correlate with reduced sperm-egg binding and fusion and in turn with reduced male fertility in Inpp5b(-/-) mice. Multiple endpoints of reproductive functions [mating trials, in vitro fertilization (IVF) assays and ADAM2 and ADAM3 cleavage] were investigated on a male-by-male basis, with pair-wise correlation analysis used to assess the relationships between these various parameters. Motile sperm from Inpp5b(-/-) mice showed significantly reduced fertilization of zona pellucida-free eggs due to reduced binding to the egg plasma membrane and subsequent fusion. Localization of a mouse sperm protein required for gamete fusion, IZUMO1, appears normal in Inpp5b-null sperm. To our surprise and differing from previous reports, we found that ADAM cleavage was only modestly impaired in numerous Inpp5b-null males and varied between individual animals. Performance in mating trials also differed from past reports. The pair-wise correlation analysis revealed that ADAM2 and ADAM3 cleavage was positively correlated, suggesting that processing of these proteins occurs by related/identical mechanisms, but otherwise, there were few correlations between the reproductive endpoints examined here. Nevertheless, this work provides detailed analysis of the Inpp5b(-/-) phenotype and also a blueprint for multivariate analysis to examine relationships between molecular characteristics and in vitro and in vivo physiological functions.
authors.affiliation: Division of Reproductive Biology, Department of Biochemistry and Molecular Biology, Johns Hopkins Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe St., Baltimore, MD 21205, USA.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Marcello
authors.forename: Janice P
authors.initials: JP
authors.lastname: Evans
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000818:Animals; D005260:Female; D000072196:Fertilins; D005298:Fertility / Q000235:genetics / Q000502:physiology*; D007136:Immunoglobulins / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008817:Mice, Mutant Strains; D015999:Multivariate Analysis; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics*; D013084:Sperm-Ovum Interactions / Q000235:genetics / Q000502:physiology*; D013094:Spermatozoa / Q000378:metabolism*
chemical_list: D007136:Immunoglobulins; C498177:Izumo1 protein, mouse; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010744:Phosphoric Monoester Hydrolases; D051722:ADAM Proteins; C000606458:Adam2 protein, mouse; C485429:Adam3 protein, mouse; D000072196:Fertilins
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