Cyclic AMP is a requisite messenger in the action of big PTTH in the prothoracic glands of pupal Manduca sexta.
Prothoracicotropic hormone (PTTH), a peptide produced by the insect brain, stimulates the prothoracic glands to secrete ecdysteroids. The big form of this peptide (25.5 kDa) has been postulated to act through cyclic AMP in larval Manduca sexta, but the role of the cyclic nucleotide in the action of PTTH in pupal glands has been less clear. Results of the present study indicate that PTTH-stimulated ecdysteroid secretion and protein phosphorylation by glands removed from pupal Manduca sexta are blocked by two inhibitors of cAMP-dependent protein kinase: Rp-cAMPS, an antagonist of cAMP binding to the regulatory subunit of the kinase, and H-89, an inhibitor of the catalytic subunit of the kinase. Further, PTTH stimulates significant accumulation of cAMP in pupal glands, although less than that previously seen in PTTH-stimulated larval glands. Cyclic AMP-dependent protein kinase is found in cytoplasmic and membrane-associated glandular subfractions, as measured by incorporation of [32P]8-N3cAMP into the regulatory subunit of the kinase. PTTH enhances cytoplasmic cAMP content and appears to increase the amount of cAMP bound to a cytoplasmic type II regulatory subunit of cAMP-dependent protein kinase. The results indicate that cAMP plays a requisite role in PTTH action in pupal glands, thus arguing in favor of a uniform mechanism of action for the peptide during Manduca development.
Smith
W A
WA
Department of Biology, Northeastern University, Boston, MA 02115, U.S.A.
Varghese
A H
AH
Healy
M S
MS
Lou
K J
KJ
D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism*; D004440:Ecdysone / Q000378:metabolism; D017952:Ganglia, Invertebrate / Q000378:metabolism*; D007301:Insect Hormones / Q000378:metabolism*; D018402:Manduca / Q000378:metabolism*; D011679:Pupa; D015398:Signal Transduction
D007301:Insect Hormones; D004440:Ecdysone; C009089:prothoracicotropic hormone; D000242:Cyclic AMP