Basolateral regulation of pH(i) in isolated snake renal proximal tubules in presence and absence of bicarbonate

William H Dantzler, Oscar K. Serrano, Diane E. Abbott, Yung Kyu Kim, Olga H. Brokl

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Intracellular pH (pH(i)) and its basolateral regulation were studied in isolated proximal-proximal and distal-proximal segments of garter snake (Thamnophis spp.) renal tubules with oil-filled lumens in HEPES-buffered and in HEPES-HCO3/--buffered media (pH 7.4 at 25°C). pH(i) was measured with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6- carboxyfluorescein (BCECF) under resting conditions and in response to NH4Cl pulse. Resting pH(i) (~7.1-7.2) and its response to and rate of recovery (dpH(i)/dt) from an NH4Cl pulse were not affected by the presence or absence of HCO3/- in either segment. Rate of recovery was depressed by Na+ removal in distal-proximal segments only and only in HEPES buffer. It was not affected by removal of Cl- or of both Na+ and Cl- or by reduction in membrane potential through addition of Ba2+ (5 mM) or high K+ (75 mM) in either segment in either HEPES or HEPES-HCO3/- buffer. The Na+/H+ exchange inhibitor ethylisopropylamiloride (EIPA) (100 μM) and the anion exchange inhibitor DIDS (100 μM) reduced dpH(i)/dt in the distal-proximal segments only and only in HEPES-HCO3 buffer. The H+-ATPase inhibitor bafilomycin (1 μM), H+-K+-ATPase and K+/NH4/+ exchange inhibitor Schering 28080 (10-100 μM), organic cation efflux inhibitor tetrapentylammonium (25 μM-20 mM), and K+ channel blocker tetraethylammonium (20 mM) had no effect on dpH(i)/dt in either segment. These data do not clearly support basolateral regulation of pH(i) in snake proximal renal tubules by commonly recognized Na+-dependent or Na+- independent acid or base transporters.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Regulatory Integrative and Comparative Physiology
Volume276
Issue number6 45-6
StatePublished - Jun 1999

Fingerprint

Proximal Kidney Tubule
Snakes
HEPES
Bicarbonates
Colubridae
Buffers
Proton-Translocating ATPases
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Tetraethylammonium
Fluorescent Dyes
Membrane Potentials
Anions
Cations
Oils
Kidney
Acids

Keywords

  • Ammonium chloride pulse
  • Bafilomycin
  • Buffers
  • Garter snakes
  • Schering 28080
  • Thamnophis spp.

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Basolateral regulation of pH(i) in isolated snake renal proximal tubules in presence and absence of bicarbonate. / Dantzler, William H; Serrano, Oscar K.; Abbott, Diane E.; Kim, Yung Kyu; Brokl, Olga H.

In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology, Vol. 276, No. 6 45-6, 06.1999.

Research output: Contribution to journalArticle

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abstract = "Intracellular pH (pH(i)) and its basolateral regulation were studied in isolated proximal-proximal and distal-proximal segments of garter snake (Thamnophis spp.) renal tubules with oil-filled lumens in HEPES-buffered and in HEPES-HCO3/--buffered media (pH 7.4 at 25°C). pH(i) was measured with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6- carboxyfluorescein (BCECF) under resting conditions and in response to NH4Cl pulse. Resting pH(i) (~7.1-7.2) and its response to and rate of recovery (dpH(i)/dt) from an NH4Cl pulse were not affected by the presence or absence of HCO3/- in either segment. Rate of recovery was depressed by Na+ removal in distal-proximal segments only and only in HEPES buffer. It was not affected by removal of Cl- or of both Na+ and Cl- or by reduction in membrane potential through addition of Ba2+ (5 mM) or high K+ (75 mM) in either segment in either HEPES or HEPES-HCO3/- buffer. The Na+/H+ exchange inhibitor ethylisopropylamiloride (EIPA) (100 μM) and the anion exchange inhibitor DIDS (100 μM) reduced dpH(i)/dt in the distal-proximal segments only and only in HEPES-HCO3 buffer. The H+-ATPase inhibitor bafilomycin (1 μM), H+-K+-ATPase and K+/NH4/+ exchange inhibitor Schering 28080 (10-100 μM), organic cation efflux inhibitor tetrapentylammonium (25 μM-20 mM), and K+ channel blocker tetraethylammonium (20 mM) had no effect on dpH(i)/dt in either segment. These data do not clearly support basolateral regulation of pH(i) in snake proximal renal tubules by commonly recognized Na+-dependent or Na+- independent acid or base transporters.",
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AU - Brokl, Olga H.

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N2 - Intracellular pH (pH(i)) and its basolateral regulation were studied in isolated proximal-proximal and distal-proximal segments of garter snake (Thamnophis spp.) renal tubules with oil-filled lumens in HEPES-buffered and in HEPES-HCO3/--buffered media (pH 7.4 at 25°C). pH(i) was measured with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6- carboxyfluorescein (BCECF) under resting conditions and in response to NH4Cl pulse. Resting pH(i) (~7.1-7.2) and its response to and rate of recovery (dpH(i)/dt) from an NH4Cl pulse were not affected by the presence or absence of HCO3/- in either segment. Rate of recovery was depressed by Na+ removal in distal-proximal segments only and only in HEPES buffer. It was not affected by removal of Cl- or of both Na+ and Cl- or by reduction in membrane potential through addition of Ba2+ (5 mM) or high K+ (75 mM) in either segment in either HEPES or HEPES-HCO3/- buffer. The Na+/H+ exchange inhibitor ethylisopropylamiloride (EIPA) (100 μM) and the anion exchange inhibitor DIDS (100 μM) reduced dpH(i)/dt in the distal-proximal segments only and only in HEPES-HCO3 buffer. The H+-ATPase inhibitor bafilomycin (1 μM), H+-K+-ATPase and K+/NH4/+ exchange inhibitor Schering 28080 (10-100 μM), organic cation efflux inhibitor tetrapentylammonium (25 μM-20 mM), and K+ channel blocker tetraethylammonium (20 mM) had no effect on dpH(i)/dt in either segment. These data do not clearly support basolateral regulation of pH(i) in snake proximal renal tubules by commonly recognized Na+-dependent or Na+- independent acid or base transporters.

AB - Intracellular pH (pH(i)) and its basolateral regulation were studied in isolated proximal-proximal and distal-proximal segments of garter snake (Thamnophis spp.) renal tubules with oil-filled lumens in HEPES-buffered and in HEPES-HCO3/--buffered media (pH 7.4 at 25°C). pH(i) was measured with the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6- carboxyfluorescein (BCECF) under resting conditions and in response to NH4Cl pulse. Resting pH(i) (~7.1-7.2) and its response to and rate of recovery (dpH(i)/dt) from an NH4Cl pulse were not affected by the presence or absence of HCO3/- in either segment. Rate of recovery was depressed by Na+ removal in distal-proximal segments only and only in HEPES buffer. It was not affected by removal of Cl- or of both Na+ and Cl- or by reduction in membrane potential through addition of Ba2+ (5 mM) or high K+ (75 mM) in either segment in either HEPES or HEPES-HCO3/- buffer. The Na+/H+ exchange inhibitor ethylisopropylamiloride (EIPA) (100 μM) and the anion exchange inhibitor DIDS (100 μM) reduced dpH(i)/dt in the distal-proximal segments only and only in HEPES-HCO3 buffer. The H+-ATPase inhibitor bafilomycin (1 μM), H+-K+-ATPase and K+/NH4/+ exchange inhibitor Schering 28080 (10-100 μM), organic cation efflux inhibitor tetrapentylammonium (25 μM-20 mM), and K+ channel blocker tetraethylammonium (20 mM) had no effect on dpH(i)/dt in either segment. These data do not clearly support basolateral regulation of pH(i) in snake proximal renal tubules by commonly recognized Na+-dependent or Na+- independent acid or base transporters.

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