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Biology of Reproduction, Vol 51, 392-399, Copyright © 1994 by Society for the Study of Reproduction

ARTICLES

Detection of insulin-like growth factor binding protein-1 in cat implantation sites

RA Boomsma, PA Mavrogianis, AT Fazleabas, RC Jaffe and HG Verhage
Department of Biology, Trinity Christian College, Palos Heights, Illinois 60463.

This study was undertaken to determine whether insulin-like growth factor binding protein-1 (IGFBP-1) was synthesized by the cat uterus and placenta during implantation and pregnancy. Endometrial and placental tissue explants from pregnant, pseudopregnant, and ovariectomized steroid-treated cats were cultured in the presence of 35S-methionine. Culture media proteins were separated by one- dimensional (1-D) and two-dimensional (2-D) SDS-PAGE, transferred to nitrocellulose, and immunostained using a rabbit polyclonal antibody against baboon IGFBP-1 and a murine monoclonal antibody to human IGFBP- 1. The antibody cross-reacted with a protein with an M(r) = 30,000 and a pI = 5.1-5.4. Immunoreactive product was found in implantation site media from 16 days postcoitum (PC) through the end of pregnancy, and was confined to the superficial placental/junctional zone. Immunoreactivity was not detected in non-implantation site media until 7 wk PC and was never detected in serum or in media from liver, pseudopregnant endometrium, or endometrium from steroid-treated cats. Autoradiography and immunostaining of 2-D Western blots of culture media proteins demonstrated that implantation site and not non- implantation site tissue synthesized and released immunoreactive IGFBP- 1 into the culture medium. 125Insulin-like growth factor-1 (IGF-1) specifically bound to this protein on 1-D Western ligand blots. Avidin- biotin immunocytochemistry utilizing the monoclonal antibody was used to localize IGFBP-1 in paraffin sections. Specific immunostaining was observed in the surface and glandular epithelium of the non-site endometrium throughout pregnancy, with stromal cell staining being detected later.(ABSTRACT TRUNCATED AT 250 WORDS)




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Copyright © 1994 by the Society for the Study of Reproduction.