Intracellular pH and oxygen chemoreception in the cat carotid body in vitro.
To test the hypothesis that O2 chemoreception in the carotid body (CB) is mediated by cellular acidosis, we simultaneously measured responses of the chemosensory and intracellular pH (pHi) to agents that are known to change pHi and studied the effects of hypoxia and ischemia on these variables in the cat CB. The CB was perfused and superfused in vitro with a modified Tyrode's solution at 36.0 +/- 0.5 degrees C with or without CO2-HCO3- (pH 7.40) and equilibrated at a given PO2. Chemosensory discharges were recorded from the whole carotid sinus nerve. To measure pHi changes, the CB was loaded with the pH-sensitive indicator 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, and the fluorescence (excitation 420-490 nm, emission greater than 515 nm) was detected by an intensified charged coupled device camera with an epifluorescence macroscope. Boluses of Tyrode's solution (0.5 ml, free of CO2-HCO3-) containing sodium acetate or NH4Cl prolonged perfusion of acid Tyrode's solution (pH 7.20-6.50), and boluses of Tyrode's solution with CO2-HCO3- were used. A decrease of fluorescence indicated pHi turning acid, and an increase of fluorescence indicated a change in alkaline pHi. Chemosensory activity varied inversely with the fluorescence change after application of these agents. Interruption of perfusate flow or application of hypoxic perfusate resulted in large increases in chemosensory discharge without any change in the fluorescence. The results indicated that chemosensory responses to brief ischemia and hypoxia were not mediated by a fall of pHi of CB cells, whereas those to CO2 and extracellular acidity were associated with decreases in pHi.
Iturriaga
R
R
Department of Physiology, School of Medicine, University of Pennsylvania, Philadelphia 19104-6085.
Rumsey
W L
WL
Lahiri
S
S
Spergel
D
D
Wilson
D F
DF
D000818:Animals; D002545:Brain Ischemia / Q000378:metabolism; D002344:Carotid Body / Q000378:metabolism*; D002415:Cats; D002628:Chemoreceptor Cells / Q000378:metabolism*; D005260:Female; D005452:Fluoresceins; D006863:Hydrogen-Ion Concentration; D002534:Hypoxia, Brain / Q000378:metabolism; D066298:In Vitro Techniques; D007424:Intracellular Fluid / Q000378:metabolism; D008297:Male; D010100:Oxygen / Q000378:metabolism*; D010477:Perfusion
D005452:Fluoresceins; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; D010100:Oxygen