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The Journal of Neuroscience, January 5, 2005, 25(1):88-95; doi:10.1523/JNEUROSCI.3209-04.2005

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Cellular/Molecular
Pharmacological Properties of GABA_A Receptors in Rat Hypothalamic Neurons Expressing the -Subunit

Olga A. Sergeeva,¹ Nadja Andreeva,¹ Maurice Garret,² Annette Scherer,¹ and Helmut L. Haas¹

¹Department of Neurophysiology, Heinrich-Heine-Universität, D-40001 Düsseldorf, Germany, and ²Laboratoire de Neurophysiologie, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5543, Université de Bordeaux II, 33076 Bordeaux, France

The pharmacological properties and functional role of native GABA_A receptors (GABA_ARs) were investigated in rat hypothalamic neurons expressing the -subunit with the help of whole-cell patch-clamp recording and single-cell reverse transcription-PCR. Two cell groups were identified: histaminergic tuberomamillary and orexinergic/hypocretinergic neurons. Approximately 25% of histaminergic and 70% of orexinergic neurons contained mRNA encoding for the -subunit. Double-immunofluorescence staining revealed a somatic localization of this protein in these two neuronal groups. Constitutive activity, diazepam modulation, fast desensitization of maximal currents, and activation by propofol (6-98 µM) of GABA_ARs did not correlate with-subunit expression. Propofol at 3-12 µM potentiated GABA-mediated currents similarly in all neurons. However, noise variance analysis of GABA-mediated currents enhanced by propofol revealed a significant difference between -positive and -negative neurons. The former displayed no difference between control and potentiated responses, and, in the latter, noise was decreased in the presence of propofol. Spontaneous IPSCs recorded in cultured hypothalamic neurons were prolonged in the presence of propofol in all -negative neurons, whereas propofol-resistant IPSCs were recorded in -positive cells. The infrequent expression of the -subunit may be a key factor in the recently discovered central role of the tuberomamillary nucleus in anesthesia.

Key words: histamine; orexin; single-cell RT-PCR; primary cultures; propofol; patch-clamp

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Received Aug 5, 2004; revised November 4, 2004; accepted November 11, 2004.

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