Immunohistochemical and nucleic acid analysis of somatotropin receptor populations in the bovine ovary

Ovaries were analyzed for somatotropin receptor protein and mRNA through use of immunohistochemistry, solution hybridization/nuclease protection, Northern blotting, and reverse transcriptase polymerase chain reaction (RT-PCR). As indicated by immunoperoxidase staining, CL expressed immunoreactive somatotropin receptor (positive stain). Ovarian stroma, connective tissue, endothelium, and erythrocytes did not express somatotropin receptor (negative stain). Within the CL, somatotropin receptor protein was expressed primarily in large luteal cells whereas small luteal cells were negative. Most follicles (1-5 mm, after fixation) were negative for somatotropin receptor. On the basis of solution hybridization/nuclease protection, the mRNA for somatotropin receptor was found in greatest abundance in CL and large luteal cells and was nearly undetectable in small luteal cells or follicles (class 1, 3-5 mm; class 2, 6-9 mm; and class 3, > or = 10 mm). Northern blotting of mRNA for somatotropin receptor showed expression of somatotropin receptor mRNA transcripts in whole ovary (4.7 and 4.4 kb), CL (4.7 and 4.4 kb), and liver (4.4 kb); and RT-PCR amplified a single amino acid coding region for somatotropin receptor in CL and liver. In summary, somatotropin receptor (both immunoreactive protein and mRNA) is found primarily in the large luteal cell, and lesser amounts of the expressed receptor or its message are found in the follicle. Alternative sizes of mRNA for somatotropin receptor suggest novel mRNA processing in the bovine ovary.