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1 Department of Biological Information Processing, Graduate School of Electronic Science and Technology, Shizuoka University, Hamamatsu, Shizuoka 432-8011, Japan2 Department of Physiology, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 431-3192, Japan3 Institute of Physiology and Pathophysiology, Johannes Gutenberg-University, D-55099 Mainz, Germany
GABA is the principal inhibitory neurotransmitter in the mature brain, but during early postnatal development the elevated [Cl–]i in immature neocortical neurones causes GABAA receptor activation to be depolarizing. The molecular mechanisms underlying this intracellular Cl– accumulation remain controversial. Therefore, the GABA reversal potential (EGABA) or [Cl–]i in early postnatal rat neocortical neurones was measured by the gramicidin-perforated patch-clamp method, and the relative expression levels of the cation–Cl– cotransporter mRNAs (in the same cells) were examined by semiquantitative single-cell multiplex RT-PCR to look for statistical correlations with [Cl–]i. The mRNA expression levels were positively (the Cl– accumulating Na+,K+–2Cl– cotransporter NKCC1) or negatively (the Cl– extruding K+–Cl– cotransporter KCC2) correlated with [Cl–]i. NKCC1 mRNA expression was high in early postnatal days, but decreased during postnatal development, whereas KCC2 mRNA expression displayed the opposite pattern. [Cl–]i and NKCC1 mRNA expression were each higher in cortical plate (CP) neurones than in the presumably older layer V/VI pyramidal neurones in a given slice. The pharmacological effects of bumetanide on EGABA were consistent with the different expression levels of NKCC1 mRNA. These data suggest that NKCC1 may play a pivotal role in the generation of GABA-mediated depolarization in immature CP cells, while KCC2 promotes the later maturation of GABAergic inhibition in the rat neocortex.
(Received 7 February 2004;
accepted after revision 13 April 2004;
first published online 16 April 2004)
Corresponding author J. Yamada: Department of Biological Information Processing, Graduate School of Electronic Science and Technology, Shizuoka University, Hamamatsu, Shizuoka 432-8011, Japan. Email: djyamad{at}ipc.shizuoka.ac.jp
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