Reduced fertility in vitro in mice lacking the cystatin CRES (cystatin-related epididymal spermatogenic): rescue by exposure of spermatozoa to dibutyryl cAMP and isobutylmethylxanthine.
The cystatin CRES (cystatin-related epididymal spermatogenic; Cst8) is the defining member of a reproductive subgroup of family 2 cystatins of cysteine protease inhibitors and is present in the epididymis and spermatozoa, suggesting roles in sperm maturation and fertilization. To elucidate the role of CRES in reproduction, mice lacking the Cst8 gene were generated and their fertility examined. Although both male and female Cst8(-/-) mice generated offspring in vivo, spermatozoa from Cst8(-/-) mice exhibited a profound fertility defect in vitro. Compared to spermatozoa from Cst8(+/+) mice, spermatozoa from Cst8(-/-) mice were unable to undergo a progesterone-stimulated acrosome reaction and had decreased levels of protein tyrosine phosphorylation, suggesting a defect in the ability of Cst8(-/-) spermatozoa to capacitate. Incubation of Cst8(-/-) spermatozoa with dibutyryl cAMP and 3-isobutyl-1-methylxanthine rescued the fertility defect, including the capacity for sperm protein tyrosine phosphorylation. Both untreated Cst8(+/+) and Cst8(-/-) spermatozoa, however, exhibited similar increased total levels of cAMP and protein kinase A (PKA) activity throughout the capacitation time course compared to spermatozoa incubated under noncapacitating conditions. Taken together, these results suggest that mice lacking CRES may have altered local levels of cAMP/PKA activity, perhaps because of improper partitioning or tethering of these signaling molecules, or that the CRES defect does not directly involve cAMP/PKA but other signaling pathways that regulate protein tyrosine phosphorylation and capacitation.
Chau
Kim M
KM
Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock, Texas 79430, USA.
Cornwall
Gail A
GA
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology*; D000375:Aging; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology*; D002478:Cells, Cultured; D015891:Cystatins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000187:drug effects* / Q000502:physiology; D013997:Time Factors; D014970:Xanthines / Q000494:pharmacology*
D015891:Cystatins; D014970:Xanthines; C078310:Cst8 protein, mouse; D003994:Bucladesine; D015056:1-Methyl-3-isobutylxanthine
Mol Reprod Dev. 1995 May;41(1):37-46
7619504
J Neuroendocrinol. 1996 Oct;8(10):801-7
8910811
Dev Biol. 1997 Dec 15;192(2):351-63
9441673
PLoS Biol. 2006 Jan;4(1):e6
16300414
Brain. 1972;95(2):387-404
4655034
J Cell Physiol. 2009 Jan;218(1):122-34
18726997
Dev Biol. 1999 Oct 15;214(2):429-43
10525345
Mol Cell Endocrinol. 2003 Feb 28;200(1-2):1-8
12644294
J Biol Chem. 2007 Nov 9;282(45):32912-23
17855342
Front Biosci (Landmark Ed). 2009 Jan 01;14:463-74
19273078
J Biol Chem. 1991 Jul 5;266(19):12633-8
2061332
Mol Reprod Dev. 1999 Apr;52(4):445-9
10092125
J Cell Sci. 2004 Jul 15;117(Pt 16):3645-57
15252132
Neuron. 2000 Nov;28(2):385-97
11144350
Mol Pathol. 1999 Dec;52(6):332-40
10748866
Hum Mol Genet. 2004 May 15;13(10):1069-79
15044380
Nat Genet. 1998 Nov;20(3):251-8
9806543
Theriogenology. 2008 Nov;70(8):1188-96
18640708
J Neuropathol Exp Neurol. 2001 Jan;60(1):94-104
11202179
Endocrinology. 2003 Mar;144(3):901-8
12586766
Science. 2009 Jul 17;325(5938):328-32
19541956
Arch Biochem Biophys. 2009 May 1;485(1):72-81
19217882
Nat Genet. 2007 Dec;39(12):1440-2
18026100
J Androl. 2011 Jul-Aug;32(4):444-63
21051588
Biol Reprod. 2008 Sep;79(3):421-31
18448843
Mol Endocrinol. 1992 Oct;6(10):1653-64
1280328
Soc Reprod Fertil Suppl. 2007;65:245-59
17644966
Biol Reprod. 1999 Jun;60(6):1542-52
10330117
EMBO J. 2001 Sep 3;20(17):4774-81
11532941
J Pharmacol Exp Ther. 1996 Oct;279(1):114-9
8858983
Biol Reprod. 2009 Nov;81(5):939-47
19605784
Genes Cells. 2008 Oct;13(10):1001-13
18754795
Neurobiol Aging. 2004 Sep;25(8):1033-43
15212828