Sensitivity of Xenopus oocytes to changes in extracellular pH: possible relevance to proposed expression of atypical mammalian GABAB receptors.
Electrical recordings were made to characterize the sensitivity of Xenopus oocytes to changes in extracellular pH, and to determine whether rat cerebral cortex poly(A)+ RNA (mRNA) expressed GABAB receptors with atypical electrical properties. All oocytes showed some sensitivity to changes in pH, and those from a small fraction (< 10%) of frogs were found to be highly responsive to acidification of bathing Ringer. In these oocytes, reduction in extracellular pH elicited membrane current responses with two components: (1) Smooth, maintained currents, primarily associated with a decrease in K+ conductance. (2) Oscillatory Cl- currents, elicited through activation of the phosphoinositide/Ca2+ messenger pathway. Oocytes with highest levels of sensitivity responded to decreases as low as 0.1 pH unit (Ringer pH 7.0 lowered to pH 6.9). Rat cortex mRNA consistently showed strong expression of membrane current responses mediated by GABAA, glutamate, kainate, serotonin and acetylcholine (muscarinic) receptors, together with responses mediated by a variety of neuropeptide receptors. In these oocytes, enantiomers of the GABAB receptor agonist baclofen, at concentrations ranging between 1 microM and 10 mM (pH 7.0), activated no significant membrane current responses. However, at concentrations > 0.1 mM hydrochloride salts of baclofen caused appreciable acidification of Ringer solutions; for example, 1 mM baclofen lowered pH from 7.0 to 6.0. Thus, when assaying oocytes with high sensitivity to pH, failure to make the necessary re-adjustment could result in apparent baclofen responses that, in reality, are simply due to pH effects alone.(ABSTRACT TRUNCATED AT 250 WORDS)
Woodward
R M
RM
Department of Psychobiology, University of California, Irvine 92717-4550.
Miledi
R
R
D000143:Acids / Q000494:pharmacology*; D000818:Animals; D016477:Artifacts; D001418:Baclofen / Q000494:pharmacology*; D002118:Calcium / Q000502:physiology; D005260:Female; D006863:Hydrogen-Ion Concentration; D008322:Mammals / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects; D009865:Oocytes / Q000737:chemistry* / Q000187:drug effects; D010716:Phosphatidylinositols / Q000502:physiology; D011963:Receptors, GABA-A / Q000032:analysis*; D012680:Sensitivity and Specificity; D014981:Xenopus
D000143:Acids; D010716:Phosphatidylinositols; D011963:Receptors, GABA-A; D001418:Baclofen; D002118:Calcium