Newly synthesized catalytic and regulatory components of adenylate cyclase are expressed in neurites of cultured sympathetic neurons.
Forskolin- and guanine nucleotide-stimulated adenylate cyclase activities were measured in microdissected sections of neurites from small explants and in dispersed cell cultures of sympathetic ganglion neurons to determine whether a competent system for regulated formation of cAMP, consisting of both catalytic units of adenylate cyclase and regulatory GTP binding proteins, is synthesized during neurite outgrowth and where it is distributed in the neuron. An increase in both guanine nucleotide- and forskolin-dependent activity of adenylate cyclase occurred concomitantly with neurite outgrowth and was directly proportional to neurite length. Separate analysis of adenylate cyclase activity in explant cell bodies or neurites showed that the increased activity was localized entirely in the neurites, while activity in the cell bodies remained virtually constant during growth. Concentric sections of neurites of approximately 500 microns width, which contained similar volumes of neurites as determined with the indicator BCECF (Rink et al., 1982), produced similar levels of cAMP, indicating an even distribution of adenylate cyclase in the neurites. Cell bodies, when stimulated by GTP gamma S, produced 236 +/- 46 attomol cAMP/min (30 degrees C)/cell body and an additional 52.6 +/- 20 attomol cAMP/min (30 degrees C)/neuron were produced with each day of neurite growth (approximately 400 microns). Assuming a turnover number of 2000 min-1, cell bodies and neurites were calculated to contain similar densities of catalytic unit molecules on their surface (9-28 molecules/micron 2). An abundant GTP binding protein, detected by ADP-ribosylation with pertussis toxin, was also widely distributed in the neuron.(ABSTRACT TRUNCATED AT 250 WORDS)
D000262:Adenylyl Cyclases / Q000096:biosynthesis*; D000818:Animals; D001369:Axons / Q000378:metabolism; D002478:Cells, Cultured; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005728:Ganglia, Sympathetic / Q000378:metabolism*; D051381:Rats; D014415:Tunicamycin / Q000494:pharmacology; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D014415:Tunicamycin; D014446:Tyrosine 3-Monooxygenase; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases