Multifactorial regulation of pituitary adenylate cyclase-activating polypeptide (PACAP)-induced production of cyclic AMP in ATT-20 corticotrophs: major involvement of Rolipram-sensitive and insensitive phosphodiesterases.
Cyclic nucleotide phosphodiesterases (PDEs) appear to play a major role in the modulation of cellular accumulations of cAMP/cGMP and hence the magnitude of the cell response to a hormone signal. These enzymes are present in cells as multiple isoforms and lie under control of various protein kinases. Because PACAP, unlike corticotropin-releasing factor (CRF), may stimulate a dual signalling pathway in pituitary cells (activating both adenylyl cyclase and phospholipase C), we used AtT-20 corticotrophs and primary cultures of rat pituitary cells to study the effect and possible differential influence of these peptides on cAMP formation. Time-course analysis indicated that, both in the absence and the presence of Rolipram (a selective type IV PDE inhibitor), PACAP stimulated a rapid and short-lived accumulation of cAMP in tumor corticotrophs, while in the presence of the non-selective inhibitor IBMX, the peptide produced a sustained high plateau level of second messenger (10 times the level generated with Rolipram at 20 min). On the contrary, when exposed to CRF, cAMP production augmented in parallel, irrespective of whether Rolipram or IBMX were present. The differential effects of the PDE inhibitors were seen with PACAP concentrations ranging from 0.1 to 100 nM, and could also be demonstrated in primary cultures of pituitary cells. Co-incubation of AtT-20 cells with Rolipram along with inhibitors of type I (but not of type III) PDEs, enhanced cAMP formation elicited by PACAP to a level significantly higher than that induced by CRF.(ABSTRACT TRUNCATED AT 250 WORDS)
Koch
B
B
Institut de Physiologie et Chimie Biologique, CNRS URA 1446, Strasbourg, France.
Lutz-Bucher
B
B
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000262:Adenylyl Cyclases / Q000378:metabolism; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D009479:Neuropeptides / Q000494:pharmacology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011760:Pyrrolidinones / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D020889:Rolipram; D019311:Staurosporine
C495403:Adcyap1 protein, rat; D000470:Alkaloids; D004791:Enzyme Inhibitors; D009479:Neuropeptides; D010726:Phosphodiesterase Inhibitors; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D011760:Pyrrolidinones; D003346:Corticotropin-Releasing Hormone; D000242:Cyclic AMP; D011493:Protein Kinase C; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D000262:Adenylyl Cyclases; D019311:Staurosporine; D020889:Rolipram; D015056:1-Methyl-3-isobutylxanthine