Regulation of functional expression of the electrogenic sodium bicarbonate cotransporter 1, NBCe1 (SLC4A4), in mouse astrocytes

Glia
Magdalena Schrödl-HäußelEleni Roussa

Abstract

The electrogenic sodium bicarbonate cotransporter NBCe1 (SLC4A4) is expressed in many cell types and is a major regulator of intracellular, and extracellular pH. In astrocytes, membrane depolarization leads to intracellular alkalinization through the activation of NBCe1. However, the molecular mechanisms regulating functional expression of NBCe1 in astrocytes are largely unknown. Astrocytes also express voltage-dependent K(+) channels that are activated after depolarization and are sensitive to the K(+) blocker 4-aminopyridine (4AP). Using acute hippocampal slices and primary hippocampal and cortical astrocyte cultures, we have investigated the role of 4AP for the regulation of NBCe1 and elucidated the underlying signaling pathways by quantitative RT-PCR, immunoblotting, biotinylation of surface proteins, immunofluorescence, and intracellular H(+) recording using the H(+) -sensitive dye 2',7'-bis-(carboxyethyl)-5-(and-6)-carboxyfluorescein. The results show significant upregulation of NBCe1 transcript, protein, and surface expression after the application of 4AP in both hippocampal slices and astrocyte cultures, effects that were suppressed after the inhibition of c-jun N-terminal kinase (JNK), proto-oncogene tyrosine-protein k...Continue Reading

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