The V‐ATPase is expressed in the choroid plexus and mediates cAMP‐induced intracellular pH alterations

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• 作者：Henriette L. Christensen ; Teodor G. Păunescu ; Vladimir Matchkov ; Dagne Barbuskaite ; Dennis Brown ; Helle H. Damkier ; Jeppe Praetorius
• 刊名：Physiological Reports
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：5
• 期：1
• 全文大小：3.0MB
• ISSN：2051-817X

文摘

The cerebrospinal fluid (CSF) pH influences brain interstitial pH and, therefore, brain function. We hypothesized that the choroid plexus epithelium (CPE) expresses the vacuolar H⁺-ATPase (V-ATPase) as an acid extrusion mechanism in the luminal membrane to counteract detrimental elevations in CSF pH. The expression of mRNA corresponding to several V-ATPase subunits was demonstrated by RT-PCR analysis of CPE cells (CPECs) isolated by fluorescence-activated cell sorting. Immunofluorescence and electron microscopy localized the V-ATPase primarily in intracellular vesicles with only a minor fraction in the luminal microvillus area. The vesicles did not translocate to the luminal membrane in two in vivo models of hypocapnia-induced alkalosis. The Na⁺-independent intracellular pH (pH_i) recovery from acidification was studied in freshly isolated clusters of CPECs. At extracellular pH (pH_o) 7.4, the cells failed to display significant concanamycin A-sensitive pH_i recovery (i.e., V-ATPase activity). The recovery rate in the absence of Na⁺ amounted to <10% of the pH_i recovery rate observed in the presence of Na⁺. Recovery of pH_i was faster at pH_o 7.8 and was abolished at pH_o 7.0. The concanamycin A-sensitive pH_i recovery was stimulated by cAMP at pH 7.4 in vitro, but intraventricular infusion of the membrane-permeant cAMP analog 8-CPT-cAMP did not result in trafficking of the V-ATPase. In conclusion, we find evidence for the expression of a minor fraction of V-ATPase in the luminal membrane of CPECs. This fraction does not contribute to enhanced acid extrusion at high extracellular pH, but seems to be activated by cAMP in a trafficking-independent manner.