Isoforms of Na,K-ATPase alpha and beta subunits in the rat cerebellum and in granule cell cultures.

There are multiple isoforms of the Na,K-ATPase in the nervous system, three isoforms of the alpha subunit, and at least two of the beta subunit. The alpha subunit is the catalytic subunit. The beta subunit has several roles. It is required for enzyme assembly, it has been implicated in neuron-glia adhesion, and the experimental exchange of beta subunit isoforms modifies enzyme kinetics, implying that it affects functional properties. Here we describe the specificities of antibodies against the Na,K-ATPase beta subunit isoforms beta1 and beta2. These antibodies, along with antibodies against the alpha subunit isoforms, were used to stain sections of the rat cerebellum and cultures of cerebellar granule cells to ascertain expression and subcellular distribution in identifiable cells. Comparison of alpha and beta isoform distribution with double-label staining demonstrated that there was no preferential association of particular alpha subunits with particular beta subunits, nor was there an association with excitatory or inhibitory neurotransmission modes. Isoform composition differences were seen when Purkinje, basket, and granule cells were compared. Whether beta1 and beta2 are specific for neurons and glia, respectively, has been controversial, but expression of both beta subunit types was seen here in granule cells. In rat cerebellar astrocytes, in sections and in culture, alpha2 expression was prominent, yet the expression of either beta subunit was low in comparison. The complexity of Na,K-ATPase isoform distribution underscores the subtlety of its regulation and physiological role in excitable cells.