Cyclic AMP regulates processing of neuropeptide precursor in bag cell neurons of Aplysia.
The stimulation of a prolonged afterdischarge of action potentials in the bag cell neurons of Aplysia is accompanied by an elevation of cAMP levels in these cells. Such a discharge causes the release of egg-laying hormone (ELH) and several other neuroactive peptides, which are derived from a 32-kDa protein prohormone. We have examined the relationship between the elevation of cAMP levels and the processing of the 32-kDa ELH prohormone. The ELH prohormone was radiolabeled in bag cell clusters by incubation of abdominal ganglia in [3H]leucine and identified on SDS-PAGE by its specific localization to bag cell neurons and its immunoreactivity with antisera to ELH. After labeling the prohormone, further incorporation of [3H]leucine was blocked using either the protein synthesis inhibitor anisomycin or an excess of unlabeled leucine. The stimulation of an afterdischarge, or treatment of cells with the adenylate cyclase activator forskolin or a membrane permeant cAMP analog, resulted in the loss of radiolabeled 32-kDa ELH prohormone relative to that in control clusters. In the presence of tetrodotoxin (TTX), which prevents discharges and stimulation-evoked secretion in the bag cell neurons, forskolin also caused the depletion of labeled ELH prohormone, suggesting that secretion per se is not likely to be required for this effect. The decrease in intensity of the 32-kDa band was accompanied by an increase in a 29-kDa band within the somata. Occasionally, an increase in a group of faint bands with approximate Mr of 26-kDa was observed. Comparative peptide mapping indicated that the 29-kDa protein is likely to be derived from the 32-kDa ELH prohormone. Our findings suggest that elevations of cAMP accelerate and possibly alter the pattern of, processing of the 32-kDa ELH prohormone.
Azhderian
E M
EM
Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510.
Kaczmarek
L K
LK
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D001048:Aplysia / Q000187:drug effects / Q000502:physiology*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology*; D007447:Invertebrate Hormones / Q000096:biosynthesis*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013779:Tetrodotoxin / Q000494:pharmacology; D013806:Theophylline / Q000494:pharmacology
D007447:Invertebrate Hormones; D011498:Protein Precursors; C026880:egg-laying hormone, Mollusca; D005576:Colforsin; D013779:Tetrodotoxin; D013806:Theophylline; D000242:Cyclic AMP