Detection of Insulin-Like Growth Factor Binding Protein-1 in Cat Implantation Sites1
| Autor: | Randal C. Jaffe, Patricia A. Mavrogianis, Harold G. Verhage, Asgerally T. Fazleabas, Robert A. Boomsma |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
medicine.medical_specialty
biology Growth factor medicine.medical_treatment Immunocytochemistry Trophoblast Cell Biology General Medicine Endometrium Molecular biology Insulin-like growth factor-binding protein Endocrinology medicine.anatomical_structure Syncytiotrophoblast Reproductive Medicine Placenta Internal medicine medicine biology.protein Immunostaining |
| Zdroj: | Biology of Reproduction. 51:392-399 |
| ISSN: | 1529-7268 0006-3363 |
| DOI: | 10.1095/biolreprod51.3.392 |
| Popis: | This study was undertaken to determine whether insulin-like growth factor binding protein-i (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 5S-methionine. Culture media proteins were separated by one-dimensional ( -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, = 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. "'5Insulin-like growth factor-I (IGF-I) 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 nonsite endometrium throughout pregnancy, with stromal cell staining being detected later. The placental labyrinth had widespread specific immunostaining, especially in the syncytiotrophoblast and maternal giant cells after 4 wk; after 9 wk, immunostaining could be detected only in the giant cells. All cells in the junctional zone and the deep glandular region of the implantation site contained IGFBP-1 staining. The synthesis of IGFBP-1 and its release into culture medium appears to be dependent on the process of implantation in the cat and may play an autocrine/paracrine role in the control of trophoblast growth and invasion. |
| Databáze: | OpenAIRE |
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