Uptake and ultrastructural localization of a [125I] growth hormone releasing factor agonist in male rat pituitary gland: evidence for internalization.
GRF was isolated from a human tumor of the pancreas and characterized. GRF stimulates the in vivo and in vitro secretion of GH. The present study was designed to find out whether human (h) GRF agonist could be internalized and to determine the subcellular localization of internalized peptide in somatotrophs. Autoradiography was performed on rat anterior pituitary glands removed at specific time intervals (2-60 min) after iv injection of monoradioiodinated [125I] (His1,Nle27) hGRF (1-32) NH2. Administration of an excess of unlabeled hGRF agonist along with the radioiodinated hormone prevented the uptake, indicating the specificity of the reaction. At the ultrastructural level only the somatotrophs appeared to contain silver grains. The main effect of hGRF agonist injection on the cytological aspect of the somatotrophs was a decrease in the area occupied by secretory granules, accompanied inversely, by an increase in that of the Golgi complex. The time course study in somatotrophs showed that five compartments (plasma membrane, secretory granules, cytoplasmic matrix, nuclear membrane, and lysosomes) have distinct marked labeling patterns. Plasma membrane, secretory granules, and nuclear membrane were labeled throughout the time course studied (2-60 min after injection). Cytoplasmic matrix was labeled 5 min post injection and lysosomes 15 and 30 min after injection. The Golgi complex, mitochondria, rough endoplasmic reticulum, and nucleus matrix were not labeled. The findings show the cellular specificity of GRF uptake by somatotrophs and the internalization process from the plasma membrane to the intracellular organelles (secretory granules, lysosomes, and nuclear membrane). Labeling of the secretory granule compartment suggests that granules may bind and protect internalized peptide from lysosomal degradation. The appearance of label on the nuclear membrane suggests that GRF may have effects on the translocation of messenger RNA from nucleus to cytoplasm in somatotrophs.
Morel
G
G
CNRS URA 1454, Department of Histology, Lyon I University, Lyon-Sud Medical School, Oullins, France.
D000818:Animals; D001345:Autoradiography; D001692:Biological Transport; D013007:Growth Hormone-Releasing Hormone / Q000031:analogs & derivatives* / Q000378:metabolism; D007457:Iodine Radioisotopes; D007700:Kinetics; D008297:Male; D008854:Microscopy, Electron; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors
D007457:Iodine Radioisotopes; D013007:Growth Hormone-Releasing Hormone; C066200:somatotropin-releasing hormone (1-32)amide, His(1),Nle(27)-