Electrophysiological properties and cholinergic responses in guinea-pig celiac ganglion neurons in primary culture.
Prevertebral neurons enzymatically dissociated from celiac ganglia of adult guinea-pigs were maintained in long-term primary culture. Cells were plated at a density of 95 +/- 15 cm-2, and intracellular electrical activity was measured between 2 and 7 weeks after dissociation. Neurite outgrowth began within 24 h of enzymatic dissociation. Cell survival dropped below 50% after more than two weeks in culture. The resting potential (-53 mV +/- 0.8), time constant (12 ms +/- 1.3), input resistance (47 M omega +/- 8.6), rheobase (0.33 nA +/- 0.02), degree of accommodation, spike amplitude (70 mV +/- 3.0), after hyperpolarization amplitude (-9.5 mV +/- 0.55), and after hyperpolarization duration (88 ms +/- 7.6) in these cells were not different from those recorded from neurons in intact celiac ganglia. A larger proportion (greater than 90%) of cells exhibited fast accommodation (phasic) in response to depolarizing current pulses. Unevoked (spontaneous) depolarizations and action potentials were observed. The cells responded to pressure ejected acetylcholine. Two types of responses consisted of an early rapid depolarization which was attenuated by hexamethonium and a later slow depolarization which was attenuated by atropine. We conclude that prevertebral neurons from guinea-pigs can be maintained in long-term primary culture, that they retain electrophysiological properties similar to intact ganglia and exhibit complex responsivity to acetylcholine.
Department of Pharmacology, College of Medicine, University of Arizona, Tucson 85724.
J S
JS
Coggan
R
R
Gruener
D L
DL
Kreulen
D000109:Acetylcholine / Q000494:pharmacology; D000200:Action Potentials; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D002470:Cell Survival; D002478:Cells, Cultured; D005260:Female; D005728:Ganglia, Sympathetic / Q000166:cytology*; D006168:Guinea Pigs; D018738:Hexamethonium; D006584:Hexamethonium Compounds / Q000494:pharmacology; D008564:Membrane Potentials; D009474:Neurons / Q000187:drug effects / Q000502:physiology*
D006584:Hexamethonium Compounds; D018738:Hexamethonium; D001285:Atropine; D000109:Acetylcholine