Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8839103 | IBRO Reports | 2018 | 6 Pages |
Abstract
The neuropeptides orexin and melanin-concentrating hormone (MCH), as well as the neurotransmitters GABA (γ-Aminobutyric acid) and glutamate are chief modulators of the sleep-wake states in the posterior hypothalamus. To investigate co-expression of vesicular GABA transporter (VGAT, a marker of GABA neurons) and the vesicular glutamate transporter-2 (VGLUT2, a marker of glutamate neurons) in orexin and MCH neurons, we generated two transgenic mouse lines. One line selectively expressed the reporter gene EYFP in VGAT+ neurons, whereas the other line expressed reporter gene tdTomato in VGLUT2+ neurons. Co-localization between these genetic reporters and orexin or MCH immunofluorescent tags was determined using 3D computer reconstructions of Z stacks that were acquired using a multiphoton laser confocal microscope. Our results demonstrated that MCH neurons expressed neither VGAT nor VGLUT2, suggesting MCH neurons are a separate cluster of cells from VGAT+ GABAergic neurons and VGLUT2+ glutamatergic neurons. Moreover, most orexin neurons expressed VGLUT2, indicating these neurons are glutamatergic. Our data suggested that in the posterior hypothalamus there are four major distinct groups of neurons: VGAT+, orexin+/VGLUT2+, orexin-/VGLUT2+, and MCH neurons. This study facilitated our understanding of the role of these neurotransmitters and neuropeptides in relation to sleep/wake regulation.
Keywords
GAD65glutamic acid decarboxylase-67glutamate decarboxylase 1GAD1MCHRTNVGLUT2VGATSSCREMNREMGad67CeAAminobutyric acidarousalvesicular GABA transporterrapid eye movementnon-rapid eye movementSleepSomatosensory cortexreticular thalamic nucleuscentral nucleus of amygdalaMelanin concentrating hormoneGABAglutamate
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Authors
Carlos Blanco-Centurion, Emmaline Bendell, Bingyu Zou, Ying Sun, Priyattam J. Shiromani, Meng Liu,