IN-SITU GENE-EXPRESSION OF GROWTH-HORMONE (GH) RECEPTOR AND GH BINDING-PROTEIN IN ADULT MALE-RAT TISSUES

Pituitary growth hormone (GH) acts as a growth promoter in a wide range of tissues after binding to its specific GH receptor (GHR) or to a cytosolic circulating GH binding protein (GHBP). To further characterize GH target cells in the rat, in situ hybridization was used to investigate the tissue and cell distribution of mRNAs encoding GHR and GHBP, and their hepatic developmental expression was examined. Cryostat sections of adult male rat tissue were hybridized with [S-35]dATp-labeled oligonucleotide antisense probes, one directed against a specific sequence of the intracellular domain of rat GHR mRNA, the other against the hydrophilic tail of rat GHBP mRNA. Several tests were carried out to validate the in situ detection of mRNA. Co-expression of the two transcripts in liver, spleen, thymus, kidney, adrenal, skin, muscle, heart, and pituitary was autoradiographically detected. However, relative expression levels, as demonstrated by computer-assisted microdensitometry, appeared to be variable. Both transcripts showed higher levels of expression in the liver, anterior and intermediate pituitary lobes, outer kidney medulla, adrenal cortex, skin epidermis, heart and muscle, but lower levels in spleen, thymus, hypodermis, adrenal medulla and posterior pituitary lobe. As a physiological control, hepatic levels of expression were examined during development, and the two forms of mRNA were found to be present at low levels in fetal liver, increasing considerably after birth. These results permit the identification in the adult male rat of cells that might be directly responsive to GH, and demonstrate the differential expression of rGHR and rGHBP transcripts.