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1 Neurosurgery, Yale University, New Haven, CT, USA
* To whom correspondence should be addressed. E-mail: anthony.vandenpol{at}yale.edu.
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 MCH promoter driving GFP or dsRed in CNS cultures to identify live MCH neurons; all neurons expressing the reporter gene showed MCH immunoreactivity, indicating selective expression. MCH neurons have a resting membrane potential of -57.5±0.6 mV, a linear current voltage relation, and a mean input resistance of 1013M
. 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 and MCH showed no direct effect on membrane potential, input resistance, action potential width, or afterhyperpolarization potential, but inhibited voltage-dependent calcium channels in MCH neurons, indicating that MCH neurons expressed both MCH and NPY receptors.
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