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Ouabain binding kinetics and FXYD7 expression in astrocytes and neurons in primary cultures: implications for cellular contributions to extracellular K+ homeostasis?

Published online by Cambridge University Press:  26 February 2010

Liang Peng ,
Rong Huang ,
Shiquen Zhang  and
Leif Hertz
Liang Peng
Affiliation:
Department of Clinical Pharmacology, China Medical University, Shenyang, P.R. China
Rong Huang
Affiliation:
Department of Pharmacology, School of Medicine, University of Saskatchewan, Saskatoon, Canada
Shiquen Zhang
Affiliation:
Department of Clinical Pharmacology, China Medical University, Shenyang, P.R. China
Leif Hertz*
Affiliation:
Department of Clinical Pharmacology, China Medical University, Shenyang, P.R. China Department of Pharmacology, School of Medicine, University of Saskatchewan, Saskatoon, Canada
*
Correspondence should be addressed to: Leif Hertz, R.R. 2, Box 245 (538 Skene Road) Gilmour, ON K0L 1W0 Canada phone: 1 613 474 0537 fax: 1 613 474 0538 email: [email protected]
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Abstract

Although Na+,K+-ATPase-mediated K+ uptake into astrocytes plays a major role in re-establishing resting extracellular K+ following neuronal excitation little information is available about astrocytic Na+,K+-ATPase function, let alone mechanisms returning K+ to neurons. The catalytic units of the Na+,K+-ATPase are the astrocyte-specific α2, the neuron-specific α3 and the ubiquitously expressed α1. In the present work, Bmax and KD values for α1, α2 and α3 subunits were computed in cultured cerebro-cortical mouse astrocytes and cerebellar granule neurons by non-linear regression as high-affinity (α2, α3) and low-affinity (α1) [3H]ouabain binding sites, which stoichiometrically equal transporter sites. Cellular expression was also determined of the brain- and α1-β1 isoform-specific FDYX7, regulating Na+,K+-ATPase efficiency and K+-sensitivity. From ouabain-sensitive K+ uptake rates published by ourselves (Walz and Hertz, 1982) or others (Atterwill et al., 1985), Na+,K+-ATPase turnover was determined. Subunits α2 and α3 showed Bmax of 15–30 pmol/mg protein, with maximum turnover rates of 70–80/s. Bmax of the α1 subunit was low in neurons but very high in astrocytes (645 pmol/mg protein), where turnover rate was slow, reflecting expression of selectively expressed FXYD7, and binding was increased by K+. The role of these characteristics for K+ homeostasis are discussed.

Keywords

α1 isoform of Na+K+-ATPaseα2 isoform of Na+K+-ATPaseα3 isoform of Na+K+-ATPaseendogenous ouabainNa+K+-ATPase turnover rate
Type
Research Article
Information
Neuron Glia Biology , Volume 6 , Issue 2 , May 2010 , pp. 127 - 135
Copyright
Copyright © Cambridge University Press 2010

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