Processing and localization of ADAMTS-1 and proteolytic cleavage of versican during cumulus matrix expansion and ovulation.
ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motifs-1) is a member of the ADAMTS family of metalloproteases which, together with ADAMTS-4 and ADAMTS-5, has been shown to degrade members of the lectican family of proteoglycans. ADAMTS-1 mRNA is induced in granulosa cells of periovulatory follicles by the luteinizing hormone surge through a progesterone receptor-dependent mechanism. Female progesterone receptor knockout (PRKO) mice are infertile primarily due to ovulatory failure and lack the normal periovulatory induction of ADAMTS-1 mRNA. We therefore investigated the protein localization and function of ADAMTS-1 in ovulating ovaries. Antibodies against two specific peptide regions, the pro-domain and the metalloprotease domain of ADAMTS-1, were generated. Pro-ADAMTS-1 of 110 kDa was identified in mural granulosa cells and appears localized to cytoplasmic secretory vesicles. The mature (85-kDa pro-domain truncated) form accumulated in the extracellular matrix of the cumulus oocyte complex (COC) during the process of matrix expansion. Each form of ADAMTS-1 protein increased >10-fold after the ovulatory luteinizing hormone surge in wild-type but not PRKO mice. Versican is also localized selectively to the ovulating COC matrix and was found to be cleaved yielding a 70-kDa N-terminal fragment immunopositive for the neoepitope DPEAAE generated by ADAMTS-1 and ADAMTS-4 protease activity. This extracellular processing of versican was reduced in ADAMTS-1-deficient PRKO mouse ovaries. These observations suggest that one function of ADAMTS-1 in ovulation is to cleave versican in the expanded COC matrix and that the anovulatory phenotype of PRKO mice is at least partially due to loss of this function.
Russell
Darryl L
DL
Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
Doyle
Kari M H
KM
Ochsner
Scott A
SA
Sandy
John D
JD
Richards
JoAnne S
JS
D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; D000818:Animals; D000911:Antibodies, Monoclonal; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D019483:Disintegrins / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism*; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000378:metabolism; D005109:Extracellular Matrix / Q000201:enzymology / Q000378:metabolism*; D005260:Female; D006107:Granulosa Cells / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure*; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000032:analysis / Q000378:metabolism; D037181:Lectins, C-Type; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009865:Oocytes / Q000166:cytology; D010060:Ovulation; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000378:metabolism; D011980:Receptors, Progesterone / Q000235:genetics / Q000502:physiology; D014018:Tissue Distribution; D053675:Versicans
D000911:Antibodies, Monoclonal; D011508:Chondroitin Sulfate Proteoglycans; D019483:Disintegrins; D004792:Enzyme Precursors; D007527:Isoenzymes; D037181:Lectins, C-Type; D011980:Receptors, Progesterone; C508323:Vcan protein, mouse; D053675:Versicans; D010447:Peptide Hydrolases; D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; C486323:Adamts1 protein, mouse; D008666:Metalloendopeptidases