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The Journal of Neuroscience, September 15, 2000, 20(18):6839-6848

Na/HCO₃ Cotransporters in Rat Brain: Expression in Glia, Neurons, and Choroid Plexus

Bernhard M. Schmitt¹, Urs V. Berger³, Robert M. Douglas², Mark O. Bevensee¹, Matthias A. Hediger³, Gabriel G. Haddad^{1, 2}, and Walter F. Boron¹

Departments of ¹ Cellular and Molecular Physiology and ² Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06520, and ³ Harvard Institute of Medicine, Boston, Massachusetts 02115

We studied the expression and distribution of Na/HCO₃ cotransporters in rat brain using polynucleotide probes and polyclonal antibodies derived from the electrogenic rat kidney Na/HCO₃ cotransporter (rkNBC). In whole brain, we observed a single mRNA (~7.5 kb) by Northern hybridization and a major ~130 kDa protein by immunoblotting with a polyclonal antiserum directed against the C terminus of rkNBC. NBC mRNA and protein were present in cortex, brainstem-diencephalon, and cerebellum. In situ hybridization revealed NBC mRNA expression throughout the CNS, with particularly high levels in olfactory bulb, hippocampal dentate gyrus, and cerebellum. NBC mRNA was present in glial cells (e.g., Bergmann glia of cerebellum and hippocampal astrocytes) and neurons (e.g., granule cells of dentate gyrus and neurons of cortex or striatum). Double hybridization of mRNA encoding NBC and glutamate transporter 1 (glial marker) confirmed that both glia and neurons express NBC. Indirect immunofluorescence microscopy demonstrated NBC protein throughout the CNS, particularly in hippocampus and cerebellum. Although NBC mRNA was restricted to cell bodies, NBC protein was distributed diffusely, compatible with a localization in cell processes and perhaps cell bodies. Double labeling with glial fibrillary acidic protein (astrocytic marker), microtubule-associated protein 2 (neuronal marker), or 2',3'-cyclic mononucleotide 3'-phosphodiesterase (oligodendrocytic marker) demonstrated expression of NBC protein in specific subpopulations of both glia and neurons. Moreover, NBC protein was present in both cultured hippocampal astrocytes and cortical neurons. NBC mRNA and protein were also present in epithelial cells of choroid plexus, ependyma, and meninges. Our results are thus consistent with multiple novel roles for Na/HCO₃ cotransport in CNS physiology.

Key words: NBC; astrocytes; Northern hybridization; in situ hybridization; immunoblotting; indirect immunofluorescence microscopy

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Copyright © 2000 Society for Neuroscience  0270-6474/00/20186839-10$05.00/0

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