Na+/H+ exchanger (NHE) in the basolateral membrane is encoded by NHE-1 mRNA in LLC-PK1 clone 4 cells.
Several isoforms of Na+/H+ exchanger (NHE-1-5) have been identified. LLC-PK1 clone 4 (CL4) expresses the amiloride-sensitive type of NHE predominantly in the basolateral membrane, which is believed to be NHE-1. It is not clear whether CL4 expresses NHE in the apical membrane and which side of NHE is encoded by the NHE-1 mRNA. Using acidified CL4 cells on the filter membrane, we examined Na(+)-dependent pH recovery of the apical and basolateral membranes separately. Na+ applied to the apical membrane recovered cell pH. Na(+)-dependent pH recovery in the apical membrane was not inhibited by SITS, DIDS, or contralateral amiloride. Li+ but not K+, chol+, or NMG+ could replace Na+. These data are consistent with the presence of NHE in the apical membrane. Transfection with an antisense oligonucleotide corresponding to the 5' terminal site of NHE-1 cDNA of CL4 decreased NHE activity in the basolateral membrane but not in the apical membrane. We conclude that CL4 expresses NHE activities in both apical and basolateral membranes and that NHE-1 mRNA encodes NHE only in the basolateral membrane.
Hanai
J I
JI
First Department of Internal Medicine, University of Tokyo, Japan.
Takaichi
K
K
Miyajima
Y
Y
Fujita
T
T
Kurokawa
K
K
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D016764:Cell Polarity; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D018374:LLC-PK1 Cells; D012333:RNA, Messenger / Q000032:analysis*; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics / Q000378:metabolism*; D013552:Swine
D012333:RNA, Messenger; D017923:Sodium-Hydrogen Exchangers; C069139:growth factor-activatable Na-H exchanger NHE-1