Imidazole binding reagent diethyl pyrocarbonate (DEPC) inhibits avian intrapulmonary chemoreceptor discharge in vivo.
Data indicate that avian intrapulmonary chemoreceptors (IPC) transduce CO2 stimuli by sensing the products of CO2 hydration, [H+] and [HCO3-]. The alphastat regulation hypothesis of physiological pH sensitivity suggests that proteins sense [H+] through changes in the ionization state of imidazole groups (alphaIm). To test whether imidazole is involved with IPC CO2 sensitivity, we administered diethyl pyrocarbonate (DEPC) intravenously while recording from IPC exposed to varying levels of inspired CO2. At physiological pH, DEPC converts pH sensitive imidazole groups to pH-insensitive N-carbethoxyhistidyl residues. Single cell extracellular neural recordings were made from vagal filaments in anesthetized, unidirectionally ventilated Anas platyrhynchos. Without DEPC, IPC discharge rate was inversely proportional to inspired CO2 with characteristic dynamic responses to rapid CO2 alterations (n = 10). After DEPC treatment (> or = 15 mM), mean sensitivity of IPC discharge to static inspired CO2 levels was decreased 75% (P < 0.05), and mean peak dynamic IPC discharge rate was decreased 80% (P < 0.05). Additionally, we tested whether DEPC might alter IPC discharge by binding imidazole groups in the enzyme carbonic anhydrase (CA), but we found no effect on CA catalytic rate. We conclude that DEPC inhibits IPC CO2 signal transduction by modifying imidazole groups on acid-sensitive proteins other than CA, possibly membrane acid-base exchangers or ion channels. These data support the alphastat regulation hypothesis in IPC CO2 respiratory chemoreception and suggests a more direct link between H+ and membrane excitability.
Pilarski
Jason Q
JQ
Department of Biological Sciences, Northern Arizona University, Franklin and Beaver Streets, Flagstaff, Arizona 86011-5640, USA. jqp2@dana.ucc.nau.edu
Hempleman
Steven C
SC
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D002245:Carbon Dioxide / Q000008:administration & dosage; D002256:Carbonic Anhydrases / Q000097:blood; D002628:Chemoreceptor Cells / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004372:Ducks; D008168:Lung / Q000294:innervation* / Q000378:metabolism*; D010313:Partial Pressure; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D011956:Receptors, Cell Surface
C437127:O,O-diethylphosphorylcholine iodide; D011956:Receptors, Cell Surface; D010767:Phosphorylcholine; D002245:Carbon Dioxide; D002256:Carbonic Anhydrases