Video-imaging microfluorometry identifies alpha- and beta-like cell types in Madin-Darby canine kidney monolayers.
On the basis of intracellular, accumulation of c-SNAFL-2, we have identified three cell subtypes in Madin-Darby canine kidney (MDCK) monolayers. Highly fluorescent cells (HFC) have a high intracellular pH (pHi, whereas cells with medium fluorescence (MFC) have low pHi when perfused with buffer containing 125 mM Cl-. HFC express a Cl-/HCO3- exchanger on the apical but not the basolateral membrane. MFC express a Cl-/HCO3- exchanger on the basolateral but not the apical membrane. We have termed these cells beta- and alpha-MDCK cells, respectively. Cells with low fluorescence (LFC) probably extrude c-SNAFL-2 through a monocarboxylate transporter, because p-4-(chloromercuri)phenylsulfonic acid (PCMBS), an inhibitor of this transporter, leads to homogeneous fluorescence.
Ebner
S
S
Department of Physiology, University of Munich, Germany.
Marin-Grez
M
M
D002731:4-Chloromercuribenzenesulfonate; D000818:Animals; D017920:Antiporters / Q000032:analysis* / Q000378:metabolism*; D001692:Biological Transport / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002469:Cell Separation; D027963:Chloride-Bicarbonate Antiporters; D004285:Dogs; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007685:Kidney Tubules, Collecting / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D018715:Microscopy, Video; D013439:Sulfhydryl Reagents
D017920:Antiporters; D027963:Chloride-Bicarbonate Antiporters; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013439:Sulfhydryl Reagents; C094271:seminaphthofluorescein-calcein; D002731:4-Chloromercuribenzenesulfonate