Mertani H C, Morel G
CNRS URA 1454, Lyon-Sud School of Medicine, Oullins, France.
Mol Cell Endocrinol. 1995 Mar;109(1):47-61. doi: 10.1016/0303-7207(95)03485-p.
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 [35S]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.
垂体生长激素(GH)与其特异性生长激素受体(GHR)或胞质循环生长激素结合蛋白(GHBP)结合后,在多种组织中发挥生长促进作用。为了进一步表征大鼠体内的GH靶细胞,采用原位杂交技术研究编码GHR和GHBP的mRNA的组织和细胞分布,并检测它们在肝脏中的发育表达。成年雄性大鼠组织的冰冻切片与[35S]dATP标记的寡核苷酸反义探针杂交,一个探针针对大鼠GHR mRNA细胞内结构域的特定序列,另一个针对大鼠GHBP mRNA的亲水性尾部。进行了多项测试以验证mRNA的原位检测。通过放射自显影检测到肝脏、脾脏、胸腺、肾脏、肾上腺、皮肤、肌肉、心脏和垂体中两种转录本的共表达。然而,计算机辅助显微密度测定显示,相对表达水平似乎存在差异。两种转录本在肝脏、垂体前叶和中叶、肾外髓质、肾上腺皮质、皮肤表皮、心脏和肌肉中的表达水平较高,但在脾脏、胸腺、皮下组织、肾上腺髓质和垂体后叶中的表达水平较低。作为生理对照,在发育过程中检测了肝脏中的表达水平,发现这两种形式的mRNA在胎儿肝脏中的水平较低,出生后显著增加。这些结果有助于在成年雄性大鼠中鉴定可能直接对GH有反应的细胞,并证明rGHR和rGHBP转录本的差异表达。