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Volume 17, Number 10, Issue of May 15, 1997 pp. 3488-3502
Copyright ©1997 Society for Neuroscience

Isoforms of Na,K-ATPase  and  Subunits in the Rat Cerebellum and in Granule Cell Cultures

Received Feb. 12, 1997; accepted March 4, 1997.

Liang Peng¹, Pablo Martin-Vasallo², and Kathleen J. Sweadner¹

¹ Laboratory of Membrane Biology, Neuroscience Center, Massachusetts General Hospital, Charlestown, Massachusetts 02129, and ² Laboratorio de Biologia del Desarrollo, Departamento de Bioquimica y Biologia Molecular, Universidad de La Laguna, 38206 La Laguna, Tenerife, Spain

There are multiple isoforms of the Na,K-ATPase in the nervous system, three isoforms of the  subunit, and at least two of the  subunit. The  subunit is the catalytic subunit. The  subunit has several roles. It is required for enzyme assembly, it has been implicated in neuron-glia adhesion, and the experimental exchange of  subunit isoforms modifies enzyme kinetics, implying that it affects functional properties. Here we describe the specificities of antibodies against the Na,K-ATPase  subunit isoforms 1 and 2. These antibodies, along with antibodies against the  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  and  isoform distribution with double-label staining demonstrated that there was no preferential association of particular  subunits with particular  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 1 and 2 are specific for neurons and glia, respectively, has been controversial, but expression of both  subunit types was seen here in granule cells. In rat cerebellar astrocytes, in sections and in culture, 2 expression was prominent, yet the expression of either  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.

Key words: Na,K-ATPase; cerebellum; isoform localization; ion transport; granule cell; astrocyte; Purkinje cell; basket cell

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