Distinct expression and subcellular localization patterns of Na +/HCO 3− cotransporter (SLC 4A4) variants NBCe1-A and NBCe1-B in mouse brain

Abstract

The electrogenic Na +/HCO 3 − cotransporter (NBCe1) has been identified as a key player for regulation of intracellular pH in several cell types. The present study was undertaken to determine expression and subcellular localization of the NH 2-terminal solute carrier (SLC) 4A4 variants NBCe1-A and NBCe1-B in mouse brain using variant-specific antibodies by immunohistochemistry and immunoelectron microscopy. In addition, distribution of NBCe1 variants and activity-dependent regulation was investigated in mouse embryonic day 17.5 (E17.5) hippocampal primary cultures in vitro. The results showed NBCe1-A and NBCe1-B transcript expression in the mouse olfactory bulb, cerebral cortex, hippocampus and cerebellum. NBCe1-A was predominantly expressed in Purkinje cells, granule cells of the dentate gyrus, non-pyramidal cell bodies in cerebral cortex, and in periglomerular and mitral cells in the olfactory bulb. Pyramidal neurons in cerebral cortex and apical cell dendrites in the hippocampus were stained for both NBCe1-A and NBCe1-B. Moreover, NBCe1-B was present in Bergmann glia. At the ultrastructural level, NBCe1-B was preferentially expressed in perivascular astroglial lamellae, whereas both NBCe1 NH 2-terminal variants were localized in pre- and postsynaptic compartments. Except for the olfactory bulb, NBCe1-A was always colocalized with calbindin. Treatment of E17.5 primary hippocampal cultures with KCl, showed dramatic downregulation of NBCe1-B mRNA and protein after 60 min, whereas NBCe1-A expression remained unchanged. These data demonstrate for the first time distinct cellular distribution of NBCe1 NH 2-terminal variants in mouse brain. NBCe1 may be involved in neuronal modulation, and pH regulation during neuronal activity.