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J. Biol. Chem., Vol. 265, Issue 22, 12813-12819, Aug, 1990
S Muallem and PA Loessberg
Pancreatic acini loaded with the pH-sensitive dye 2',7'-
bis(carboxyethyl)-5(6)-carboxyfluorescein were used to examine the effect
of Ca2(+)-mobilizing agonists on the activity of acid-base transporters in
these cells. In the accompanying article (Muallen, S., and Loessberg, P. A.
(1990) J. Biol. Chem. 265, 12813-12819) we showed that in
4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid (HEPES)- buffered
medium the main pHi regulatory mechanism is the Na+/H+ exchanger, a while
in HCO3(-)-buffered medium pHi is determined by the combined activities of
a Na+/H+ exchanger, a Na(+)-HCO3- cotransporter and a Cl-/HCO3- exchanger.
In this study we found that stimulation of acini with Ca2(+)-mobilizing
agonists in HEPES or HCO3(-)-buffered media is followed by an initial
acidification which is independent of any identified plasma
membrane-located acid-base transporting mechanism, and thus may represent
intracellularly produced acid. In HEPES-buffered medium there was a
subsequent large alkalinization to pHi above that in resting cells, which
could be attributed to the Na+/H+ exchanger. Measurements of the rate of
recovery from acid load indicated that the Na+/H+ exchanger was stimulated
by the agonists. In HCO3(-)-buffered medium the alkalinization observed
after the initial acidification was greatly attenuated. Examination of the
activity of each acid-base transporting mechanism in stimulated acini
showed that in HCO3(-)-buffered medium: (a) recovery from acid load in the
presence of H2-4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (H2DIDS)
(Na+/H+ exchange) was stimulated similar to that found in HEPES-buffered
medium; (b) recovery from acid load in the presence of amiloride and
acidification due to removal of external Na+ in the presence of amiloride
(HCO3- influx and efflux, respectively, by Na(+)-HCO3- cotransport) were
inhibited; and (c) HCO3- influx and efflux due to Cl- /HCO3- exchange,
which was measured by changing the Cl- or HCO3- gradients across the plasma
membrane, were stimulated. Furthermore, the rate of Cl-/HCO3- exchange in
stimulated acini was higher than the sum of H+ efflux due to Na+/H+
exchange and HCO3- influx due to Na(+)-HCO3- cotransport. Use of H2DIDS
showed that the latter accounted for the attenuated changes in pHi in
HCO3(-)-buffered medium, as much as treating the acini with H2DIDS resulted
in similar agonist-mediated pHi changes in HEPES- and HCO3(-)-buffered
media. The effect of agonists on the various acid-base transporting
mechanisms is discussed in terms of their possible role in transcellular
NaCl transport, cell volume regulation, and cell proliferation in
pancreatic acini.
Intracellular pH-regulatory mechanisms in pancreatic acinar cells. II. Regulation of H+ and HCO3- transporters by Ca2(+)-mobilizing agonists
Department of Physiology, University of Texas, Dallas 75235.
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