Binding and internalization of gold-conjugated somatostatin and growth hormone-releasing hormone in cultured rat somatotropes.
The synthetic peptides somatostatin (SRIF) and growth hormone-releasing hormone (GRH) were coupled directly to colloidal gold of different particle sizes. Both conjugates were biologically active in displacing the corresponding radiolabeled hormones from high affinity binding sites in pituitary membranes. Release of growth hormone (GH) from cultured anterior pituitary cells was modulated by both conjugates alone or in combination. Ultrastructural studies were performed with cells incubated at 4 degrees C (2 h) and 37 degrees C (2 min-2 h) with one of the labeled peptides or their combination. Somatotropes were identified by immunostaining with anti-rGH followed by protein A-ferritin, thus obtaining a triple labeling. Both hormone conjugates were internalized in different vesicles in the beginning but accumulated during longer incubation times in the same compartment. The secretory vesicles and the nucleus were not labeled by any hormone conjugate. In contrast to SRIF-gold, the uptake of GRH-gold conjugate decreased with longer incubation times. This effect could be neutralized by simultaneous incubation of the somatotropes with both regulating hormones. Hence, whereas the binding and internalization of SRIF by somatotropes do not seem to be influenced by GRH, the corresponding processes for GRH are stimulated by the presence of SRIF.
Mentlein
R
R
Anatomisches Institut Universität Kiel, Federal Republic of Germany.
Buchholz
C
C
Krisch
B
B
D000818:Animals; D002478:Cells, Cultured; D006046:Gold; D013006:Growth Hormone / Q000378:metabolism*; D013007:Growth Hormone-Releasing Hormone / Q000493:pharmacokinetics*; D008297:Male; D008854:Microscopy, Electron; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013004:Somatostatin / Q000493:pharmacokinetics*
D013004:Somatostatin; D006046:Gold; D013006:Growth Hormone; D013007:Growth Hormone-Releasing Hormone