title: Peptide modulation of ACh receptor desensitization controls neurotransmitter release from chicken sympathetic neurons.
abstract: 1. The distribution and release of substance P (SP) in embryonic chicken lumbar sympathetic ganglia was examined with the use of immunohistochemistry and radioimmunoassay, respectively. SP immunoreactivity was detected in nerve fibers surrounding individual sympathetic neurons and was released by ganglionic depolarization. 2. Effects of SP on nicotinic acetylcholine receptor (AChR) function was assayed in embryonic sympathetic neurons in vitro by whole-cell patch clamp. SP (0.1-20 microM) accelerated the rate of decay (desensitization) of ACh-induced currents. The AChR desensitization time course is biphasic and described by the sum of two exponential functions dependent on agonist concentration (time constant of the faster component, tau f = 1-2 s, and the slower time constant, tau s = 10-25 s). SP selectively decreased tau s and the contribution of the slow component to the overall rate of current decay. The effects of SP on desensitization were concentration dependent and reversible. SP slowed recovery from desensitization by 2.5-fold. 3. SP shifted the dose-response curve for ACh-induced desensitization, reducing the concentration of ACh required to produce half-maximal desensitization by approximately twofold. 4. Preapplication of SP was equivalent to SP applied together with ACh in accelerating AChR desensitization. SP did not alter the time course of currents elicited by nondesensitizing concentrations of ACh, carbamylcholine (CARB), or dimethylphenylpiperazinium (DMPP). These data suggest that AChR activation is neither necessary nor sufficient for the peptide to modulate receptor function. A kinetic model of the effects of SP on specific steps in AChR desensitization is presented. 5. SP enhanced the rate of decay of synaptic currents in sympathetic neurons innervated in vitro, decreasing the synaptic current duration by up to 80%. 6. Effects of SP on neurotransmitter release from sympathetic neurons were evaluated by measuring the release of [3H]-norepinephrine (NE). ACh and CARB stimulated NE release in a concentration- and calcium-dependent manner. SP alone had no effect on NE secretion, but the peptide inhibited NE release induced by ACh or CARB by 40-50%. 7. Although agonists specific for either nicotinic or muscarinic receptors stimulated release of NE, SP selectively inhibited the nicotinic component of transmitter secretion. Thus SP suppressed NE release induced by DMPP by up to 80% but had no effect on muscarine or depolarization-induced NE secretion. 8. Parallel studies of the modulatory effects of SP on whole-cell currents and NE secretion revealed that SP inhibition of transmitter release from sympathetic neurons is directly proportional to the extent of potentiation of AChR desensitization.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Anatomy and Cell Biology, Columbia University College of Physicians and Surgeons, New York, New York 10032.
authors.forename: D C
authors.initials: DC
authors.lastname: Valenta
authors.forename: J E
authors.initials: JE
authors.lastname: Downing
authors.forename: L W
authors.initials: LW
authors.lastname: Role
mesh_terms: D000818:Animals; D002642:Chick Embryo; D046508:Culture Techniques; D005728:Ganglia, Sympathetic / Q000502:physiology*; D007473:Ion Channels / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D009412:Nerve Fibers / Q000502:physiology; D009474:Neurons / Q000502:physiology; D009479:Neuropeptides / Q000502:physiology*; D018377:Neurotransmitter Agents / Q000502:physiology*; D009638:Norepinephrine / Q000502:physiology; D011950:Receptors, Cholinergic / Q000502:physiology*; D013004:Somatostatin / Q000502:physiology; D013373:Substance P / Q000502:physiology; D009435:Synaptic Transmission / Q000502:physiology*
chemical_list: D007473:Ion Channels; D009479:Neuropeptides; D018377:Neurotransmitter Agents; D011950:Receptors, Cholinergic; D013373:Substance P; D013004:Somatostatin; D009638:Norepinephrine