By Felix Bronner and Arnost Kleinzeller (Eds.)
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Additional info for Cellular Mechanisms of Renal Tubular Ion Transport
It is constructed by introducing a predrawn, pH-sensitive pipet into an open-tipped lead glass pipet. The pH glass is advanced until it protrudes through the lead glass, and then the two are welded together. Kostyuk and Sorokina (28) produced an electrode with similar geometry, but with a tip small enough (-1 pm diameter) to permit impalement of frog muscle fibers. Their electrodes are made by inserting pH glass tubing into the insulating glass tubing, and then pulling the concentric capillaries on a microelectrode puller.
In mouse skeletal muscle, however, the greater part of pHi regulation appears to be via Na+-H+ exchange (1). Why the system in snail neurons should be so (apparently) complicated is unclear. Perhaps it is an advantage to use the Na gradient not only to regulate pHi , but also to keep intracellular C1- lower than would be determined by a passive distribution. Some inhibitory transmitters increase postsynaptic Cl- permeability so that for efficient inhibition Ecl needs to be more negative than Em.
An axon or muscle fiber is cannulated at both ends and mounted horizontally in a chamber. The dialysis tubing, which, in the region indicated by the broken lines, is permeant t o solutes of low molecular weight, is inserted into the fiber through the cannulas. The pH-sensitive and reference microelectrodes are introduced alongside the dialysis capillary, through opposite cannulas. The cut ends of the fiber are isolated by vaseline seals from the central region, which is superfused with ASW. INTRACELLULAR pH REGULATION 13 if pH, is raised (8).