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The Journal of Neurophysiology Vol. 85 No. 6 June 2001, pp. 2563-2575
Copyright ©2001 by the American Physiological Society
1Department of Neurological Surgery, 2Department of Physiology, and 3Department of Surgery, University of Wisconsin Medical School, Madison, Wisconsin 53792
Schomberg, Stacey L.,
Gui Su,
Robert A. Haworth, and
Dandan Sun.
Stimulation of Na-K-2Cl Cotransporter in Neurons by Activation of
Non-NMDA Ionotropic Receptor and Group-I mGluRs. J. Neurophysiol. 85: 2563-2575, 2001. In a
previous study, we found that
Na+-K+-2Cl
cotransporter in immature cortical neurons was stimulated by activation
of the ionotropic N-methyl-D-aspartate (NMDA)
glutamate receptor in a Ca2+-dependent manner. In
this report, we investigated whether the Na+-K+-2Cl
cotransporter in immature cortical neurons is stimulated by non-NMDA glutamate receptor-mediated signaling pathways. Expression of the
Na+-K+-2Cl
cotransporter and metabotropic glutamate receptors (mGluR1 and 5) was
detected in cortical neurons via immunoblotting and immunofluorescence staining. Significant stimulation of cotransporter activity was observed in the presence of both
trans-(±)-1-aminocyclopentane-trans-1,3-dicarboxylic acid
(trans-ACPD) (10 µM), a metabotropic glutamate
receptor (mGluR) agonist, and (RS)-3,5-dihydroxyphenylglycine
(DHPG) (20 µM), a selective group-I mGluR agonist. Both
trans-ACPD and DHPG-mediated effects on the cotransporter
were eradicated by
bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid-AM, a Ca2+ chelator. In addition,
DHPG-induced stimulation of the cotransporter activity was inhibited in
the presence of mGluRs antagonist (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA) (1 mM) and also with selective mGluR1 antagonist
7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester
(CPCCOEt) (100 µM). A DHPG-induced rise in intracellular Ca2+ in cortical neurons was detected with
Fura-2. Moreover, DHPG-mediated stimulation of the cotransporter was
abolished by inhibition of Ca2+/CaM kinase II.
Interestingly, the cotransporter activity was increased by activation
of 
-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)
receptor. These results suggest that the
Na+-K+-2Cl
cotransporter in immature cortical neurons is stimulated by group-I mGluR- and AMPA-mediated signal transduction pathways. The effects are
dependent on a rise of intracellular
Ca2+.
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