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This Article Full Text Full Text (PDF) All Versions of this Article: 90/6/3978    most recent 00593.2003v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (17) Citing Articles via Google Scholar Google Scholar Articles by Gao, X.-B. Articles by van den Pol, A. N. Search for Related Content PubMed PubMed Citation Articles by Gao, X.-B. Articles by van den Pol, A. N.

Neurons Synthesizing Melanin-Concentrating Hormone Identified by Selective Reporter Gene Expression After Transfection In Vitro: Transmitter Responses

Department of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut 06520

Submitted 17 June 2003; accepted in final form 22 August 2003

Neurons from the lateral hypothalamus that synthesize melanin-concentrating hormone (MCH) play an important role in the regulation of energy homeostasis. Relatively little is known of the cellular physiology and transmitter responses of these neurons, in part because of the difficulty in identifying live MCH cells. Here we use a novel approach of transfection of specific gene constructs with the MCH promoter driving green fluorescent protein (GFP) or red fluorescent protein (dsRed2) in CNS cultures to identify live rat MCH neurons; all neurons expressing the reporter gene showed MCH immunoreactivity, indicating selective expression. MCH neurons had a resting membrane potential of –57.5 ± 0.6 mV, a linear current-voltage relation and a mean input resistance of 1,013 M. Long depolarizing pulses revealed significant spike frequency adaptation. Functional glutamate and GABA receptors were expressed by MCH neurons. MCH neurons were hyperpolarized by norepinephrine in the presence or absence of tetrodotoxin, suggesting direct inhibition. Orexigenic peptides neuropeptide Y (NPY) and MCH showed no direct effect on membrane potential, input resistance, action potential width, or afterhyperpolarization potential, but inhibited voltage-dependent calcium channels, indicating that MCH neurons expressed both MCH and NPY receptors.

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