Plasticity of Na,K-ATPase isoform expression in cultures of flat astrocytes: species differences in gene expression.

The Na,K-ATPase plays an active role in glial physiology, contributing to K+ uptake as well as to the Na+ gradients used by other membrane carriers. There are multiple isoforms of Na,K-ATPase alpha and beta subunits, and different combinations result in different affinities for Na+ and K+. Isoform choice should thus influence K+ and Na+ homeostasis in astrocytes. Prior studies of astrocyte Na,K-ATPase subunit composition have produced apparently conflicting results, suggesting plasticity of gene expression. Purified flat astrocytes from the cerebral cortex and cerebellum of both mouse and rat were systematically investigated here. Using antibodies specific for the alpha1, alpha2, alpha3, beta1, beta2, and beta3 subunits, isoform level was assessed with Western blots, and cellular distribution was visualized with immunofluorescence. Although alpha1 was always expressed, differences were observed in the expression of alpha2 and beta2, subunits that can be expressed in astrocytes in vivo and in coculture with neurons. In addition, abundant alpha subunit was expressed in rat astrocytes and in mouse cerebellar astrocytes without an equivalent level of any of the known beta isoforms, suggesting that an additional beta subunit important for glia is yet to be discovered. Conditions that have been shown to increase Na,K-ATPase activity in astrocyte cultures, such as dibutyryl cAMP, high extracellular K+, and glutamate, did not specifically induce missing subunits, suggesting that cellular interactions are required to alter the ion transporter phenotype.