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.
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.
Marcello
Matthew R
MR
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.
Evans
Janice P
JP
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*
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
Reprod Fertil Dev. 2009;21(2):293-303
19210920
Biochem J. 2004 Jun 1;380(Pt 2):505-14
14972029
J Cell Sci. 1995 Oct;108 ( Pt 10):3267-78
7593287
Dev Biol. 2000 Jun 15;222(2):289-95
10837118
Nature. 2005 Mar 10;434(7030):234-8
15759005
Transgenic Res. 2010 Aug;19(4):587-94
19844803
J Exp Zool. 1976 Jul;197(1):13-9
939958
Dev Biol. 1997 Nov 1;191(1):131-45
9356177
J Cell Sci. 2009 Aug 1;122(Pt 15):2741-9
19596796
Biol Reprod. 2009 May;80(5):1001-8
19129510
Mol Reprod Dev. 2009 Dec;76(12):1188-99
19658160
J Cell Biol. 1997 Apr 7;137(1):105-12
9105040
Mol Reprod Dev. 2006 Oct;73(10):1284-91
16850391
Biol Reprod. 2009 May;80(5):897-904
19144954
J Androl. 2002 May-Jun;23(3):317-25
12002430
J Androl. 2010 Jan-Feb;31(1):34-44
19875488
Front Biosci. 2008 Jan 01;13:462-76
17981561
J Biol Chem. 1995 Apr 21;270(16):9370-7
7721860
Dev Biol. 2001 May 1;233(1):204-13
11319869
Dev Biol. 2001 Dec 15;240(2):641-53
11784089
Theriogenology. 2006 Jun;65(9):1716-26
16271754
Biol Reprod. 2009 Apr;80(4):833-41
19129508
J Reprod Fertil. 1988 Mar;82(2):681-90
3283346
Methods Mol Biol. 2006;341:89-101
16799191
Biochem J. 2009 Apr 1;419(1):29-49
19272022
J Reprod Immunol. 1987 Jun;11(2):91-100
3625612
Nat Cell Biol. 2002 Oct;4 Suppl:s41-9
12479614
J Clin Invest. 1998 May 15;101(10):2042-53
9593760
Biol Reprod. 1975 Dec;13(5):513-8
1203407
Fertil Steril. 2010 Mar 1;93(4):1112-23
19342015
Biol Reprod. 2002 May;66(5):1522-30
11967219
Nature. 1992 Jul 16;358(6383):239-42
1321346
Biol Reprod. 1976 Dec;15(5):647-53
1000005