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PH regulation in horizontal cells of the skate retina



PH regulation in horizontal cells of the skate retina



Experimental Eye Research 66(4): 449-463



To examine the mechanisms by which horizontal cells regulate intracellular pH (pHi), measurements were recorded from isolated cells enzymatically dissociated from the skate retina utilizing the pH-sensitive dye BCECF. In a HCO3--containing Ringer solution, steady-state pHi was 7.32+/-0.13 (mean+/-S.D., n=70). Recovery from acidification was examined using the NH4+ prepulse technique. When NH4+ was removed from the extracellular solution, pHi dropped rapidly to approximately 0.3 pH units below the initial baseline, and then recovered at an initial rate of approximately 0.072 pH units/min. During recovery of pHi after the acid load, the removal of Na+ or the addition of amiloride from a HCO3--free extracellular solution reduced the rate of recovery by 79%+/-11% and 69%+/-14%, respectively. In the presence of DIDS, which inhibits primarily anion transport, or during the removal of Na+, the recovery from acidification was reduced by 83%+/-10% and 70%+/-11%, respectively, as compared to the control value in HCO3--containing solution. These results suggest that the skate horizontal cell possesses a Na/H exchanger as well as a Na+-and HCO3--dependent mechanism for removal of excess acid. Removal of HCO3- or Cl- from the extracellular solution had little effect on pHi, but removing external Na+ induced a marked decrease in pHi that fell at an initial rate of approximately 0.3 pH units min-1. This rate of acidification was decreased by 58%+/-19% in the presence of DIDS (500 micron) and reduced by 28%+/-13% with the addition of amiloride (2 mm). Thus, Na- and HCO3-dependent transport was about 2-fold more active than Na/H exchange during low Na+-induced acidification. The intrinsic pH-buffer capacity, determined from the pHi change induced by incremental reductions in the [NH4+] of the extracellular solution, was 24.2 mm/pH unit at the horizontal cell's resting pHi. Moreover, pHi was relatively insensitive to changes in membrane potential; in experiments under whole-cell voltage clamp (-70 mV), intracellular pH remained constant during depolarizing voltage swings to -30 mV or +30 mV, as well as during hyperpolarizing pulses to -90 or -110 mV.

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Accession: 009743763

Download citation: RISBibTeXText

PMID: 9593638

DOI: 10.1006/exer.1997.0441


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