Altered excitability of intestinal neurons in primary culture caused by acute oxidative stress.
Neurons were isolated from the intestine of guinea pigs and grown in primary culture for < or =15 days. Using conventional whole cell recording techniques, we demonstrated that the majority of neurons express a prolonged poststimulus afterhyperpolarization (slow AHP). These neurons also had large-amplitude (approximately 100 mV), broad-duration (approximately 2 ms) action potentials and generated a hyperpolarization activated inward current (Ih). Application of H2O2 (0.22-8.8 mM) hyperpolarized these neurons but not those lacking slow AHPs. The H2O2-induced hyperpolarization was followed by irreversible depolarization at higher concentrations (more than approximately 1 mM) of H2O2 while it was maintained after washout of submillimolar H2O2. The ionic mechanisms underlying the hyperpolarization included the suppression of Ih and the activation of an inwardly rectifying outward current, which was blocked by glybenclamide (25-50 microM) and TEA (30 mM). In addition, H2O2 suppressed the slow AHP and its underlying current. Internal perfusion of catalase and glutathione opposed the H2O2-mediated decrease in IsAHP. Our results indicate that acute oxidative stress has neuron- and conductance-specific actions in intestinal neurons that may underlie pathophysiological conditions.
Vogalis
Fivos
F
Department of Anatomy and Cell Biology, University of Melbourne, Parkville, Victoria 3010, Australia.
Harvey
John R
JR
D000200:Action Potentials; D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques; D000697:Central Nervous System Stimulants / Q000494:pharmacology; D003022:Clotrimazole / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D004386:Duodenum / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D020536:Enzyme Activators / Q000494:pharmacology; D005260:Female; D005905:Glyburide / Q000494:pharmacology; D006168:Guinea Pigs; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008297:Male; D008564:Membrane Potentials; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D018384:Oxidative Stress; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers / Q000494:pharmacology; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D019789:Tetraethylammonium / Q000494:pharmacology
D000697:Central Nervous System Stimulants; D020536:Enzyme Activators; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D005576:Colforsin; D002110:Caffeine; D019789:Tetraethylammonium; D006861:Hydrogen Peroxide; D003022:Clotrimazole; D005905:Glyburide; D002118:Calcium