Ultrastructural expression of prolactin receptor in rat liver

Abstract

Prolactin (PRL) is a trophic hormone which acts mainly at the plasma membrane level of hepatocyte. The mechanisms involved in the transduction of the signal after binding of PRL to its receptors are not yet well documented. In the present study we have examined the subcellular patterns of PRL receptor expression in rat liver by ultrastructural in situ hybridization and immunocytology. In situ hybridization was performed using digoxigenin-labeled oligonucleotide probes revealed by indirect immunogold reaction. The expression of both the long and short forms of PRL-receptor mRNA was readily identified in the cytoplasmic matrix, and in association with the endoplasmic reticulum, but a low expression of these forms was detected in the nucleus of hepatocyte. Moreover, this expression appeared clearly higher in female rather than in male hepatocytes. On the other hand, immunogold detection of PRL-receptor protein was performed using two monoclonal antibodies (U5 and T6), specific to the extracellular domain of the PRL-receptor. Indirect immunocytological detection confirmed the presence of PRL receptor-like immunoreactivity at the level of the plasma membrane, and in the cytoplasmic matrix associated or not with endocytotic vesicles, the endoplasmic reticulum, the peroxisomes, the Golgi complex, and the nuclei of both male and female hepatocytes. No clear difference was found between U5 and T6 mAbs, with regard to the subcellular localization. These results show the distribution of both PRL-receptor mRNA and PRL receptor protein in numerous subcellular compartments of hepatocyte, and evidence that these compartments are involved in the early stage of the PRL action.