Differential secretion of proLHRH fragments in response to [K+], prostaglandin E2 and C kinase activation.
ProLHRH contains the luteinizing hormone-releasing hormone (LHRH) decapeptide and a 56 amino acid peptide designated gonadotropin-releasing hormone-associated peptide (GAP). We studied the effects of various known secretagogues of LHRH on the in vitro release of proLHRH fragments from the median eminence (ME) and subsequently characterized these immunoreactive products according to molecular weight (MW). GAP- and LHRH-like immunoreactive (LI) materials were secreted simultaneously into the media under basal conditions. Prostaglandin E2 stimulated release of both peptides by approximately 2-fold. Both phorbol ester and [K+] stimulated release of GAP-LI by 4-fold and LHRH-LI by 9-fold over baseline levels. When materials from [K+]-stimulated media were separated according to MW by high performance size-exclusion chromatography, a single peak eluted at 1300 MW in the same position as synthetic LHRH. Two GAP-LI peaks were observed. One eluted in the void volume, while the predominant peak co-eluted with synthetic rat GAP1-56 at approximately 6500 MW. These results indicate that GAP and LHRH are co-secreted, that they are released as intact peptides, and that activation of different intracellular pathways may cause their differential secretion. These results emphasize the importance of using both in vitro and chromatographic methodologies to evaluate the changes which may occur in LHRH prohormone processing and secretion.
Valenca
M M
MM
Reproductive Neuroendrocrinology Section, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709.
Wetsel
W C
WC
Culler
M D
MD
Negro-Vilar
A
A
D000818:Animals; D015232:Dinoprostone; D004789:Enzyme Activation; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D008473:Median Eminence / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology*; D011458:Prostaglandins E / Q000494:pharmacology*; D011493:Protein Kinase C / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D011458:Prostaglandins E; D011498:Protein Precursors; C050502:gonadotropin releasing hormone associated peptide; D007987:Gonadotropin-Releasing Hormone; D011493:Protein Kinase C; D015232:Dinoprostone; D011188:Potassium