Journal of Neuroscience, Vol 7, 110-119, Copyright © 1987 by Society for Neuroscience
Newly synthesized catalytic and regulatory components of adenylate cyclase are expressed in neurites of cultured sympathetic neurons
AM Tolkovsky
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)