TY - JOUR
T1 - Intracellular pH in isolated rat renal papillary thin limbs of Henle's loop
AU - Dantzler, William H.
AU - Kim, Yung Kyu
AU - Abbott, Diane E.
AU - Serrano, Oscar K.
AU - Brokl, Olga H.
PY - 2000
Y1 - 2000
N2 - Intracellular pH (pH(i)) was measured in isolated, nonperfused and perfused rat papillary thin limbs of Henle's loops in N-2- hydroxyethylpiperazine-N'-2-ethansulfonic acid (HEPES)- or HEPES/bicarbonate- buffered medium at pH 7.4 using the pH-sensitive fluorescent dye 2',7'-bis(2- carboxyethyl)-5,6-carboxyfluorescein (BCECF). Resting pH(i) was about 6.7 in descending thin limbs (DTL) and about 6.9 in ascending thin limbs (ATL), even with a medium pH of 7.4. These values appeared to reflect the acid pH of the blood in the neighboring vasa recta found in vivo. The resting pH(i) did not differ whether or not the medium contained bicarbonate although the total buffering capacity of the tubule cells was increased in the presence of bicarbonate. In nonperfused DTL and ATL, pH(i) was further acidified following an NH4Cl pulse. The rate of recovery of pH(i) from this level to the resting pH(i) was reduced by Na+ removal from the bath in both DTL and ATL and by the addition of ethylisopropylamiloride (EIPA) to the bath in the presence of Na+ in DTL. The rate of recovery was not affected by Cl- removal from the bath or K+ (75 mM) or 4,4'-diisothiocyanostilbene-2,2'- disulfonate (DIDS) addition to the bath in either DTL or ATL. These results suggest that the common, amiloride-sensitive, basolateral Na+/H+ exchanger plays a role in the regulation of pH(i) in rat papillary DTL but that a different basolateral Na+/H+ exchanger or a luminal Na+/H+ exchanger is important in rat papillary ATL.
AB - Intracellular pH (pH(i)) was measured in isolated, nonperfused and perfused rat papillary thin limbs of Henle's loops in N-2- hydroxyethylpiperazine-N'-2-ethansulfonic acid (HEPES)- or HEPES/bicarbonate- buffered medium at pH 7.4 using the pH-sensitive fluorescent dye 2',7'-bis(2- carboxyethyl)-5,6-carboxyfluorescein (BCECF). Resting pH(i) was about 6.7 in descending thin limbs (DTL) and about 6.9 in ascending thin limbs (ATL), even with a medium pH of 7.4. These values appeared to reflect the acid pH of the blood in the neighboring vasa recta found in vivo. The resting pH(i) did not differ whether or not the medium contained bicarbonate although the total buffering capacity of the tubule cells was increased in the presence of bicarbonate. In nonperfused DTL and ATL, pH(i) was further acidified following an NH4Cl pulse. The rate of recovery of pH(i) from this level to the resting pH(i) was reduced by Na+ removal from the bath in both DTL and ATL and by the addition of ethylisopropylamiloride (EIPA) to the bath in the presence of Na+ in DTL. The rate of recovery was not affected by Cl- removal from the bath or K+ (75 mM) or 4,4'-diisothiocyanostilbene-2,2'- disulfonate (DIDS) addition to the bath in either DTL or ATL. These results suggest that the common, amiloride-sensitive, basolateral Na+/H+ exchanger plays a role in the regulation of pH(i) in rat papillary DTL but that a different basolateral Na+/H+ exchanger or a luminal Na+/H+ exchanger is important in rat papillary ATL.
KW - Ammonium chloride pulse
KW - Henle's loops
KW - Intracellular pH
KW - Na/H exchange
KW - Total buffering capacity
UR - http://www.scopus.com/inward/record.url?scp=0034059795&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034059795&partnerID=8YFLogxK
U2 - 10.1007/s004240051031
DO - 10.1007/s004240051031
M3 - Article
C2 - 10864007
AN - SCOPUS:0034059795
SN - 0031-6768
VL - 440
SP - 140
EP - 148
JO - Pflugers Archiv European Journal of Physiology
JF - Pflugers Archiv European Journal of Physiology
IS - 1
ER -