Single-Cell Gene Expression Analysis of Cholinergic Neurons in the Arcuate Nucleus of the Hypothalamus.
The cholinoceptive system in the hypothalamus, in particular in the arcuate nucleus (ARC), plays a role in regulating food intake. Neurons in the ARC contain multiple neuropeptides, amines, and neurotransmitters. To study molecular and neurochemical heterogeneity of ARC neurons, we combine single-cell qRT-PCR and single-cell whole transcriptome amplification methods to analyze expression patterns of our hand-picked 60 genes in individual neurons in the ARC. Immunohistochemical and single-cell qRT-PCR analyses show choline acetyltransferase (ChAT)-expressing neurons in the ARC. Gene expression patterns are remarkably distinct in each individual cholinergic neuron. Two-thirds of cholinergic neurons express tyrosine hydroxylase (Th) mRNA. A large subset of these Th-positive cholinergic neurons is GABAergic as they express the GABA synthesizing enzyme glutamate decarboxylase and vesicular GABA transporter transcripts. Some cholinergic neurons also express the vesicular glutamate transporter transcript gene. POMC and POMC-processing enzyme transcripts are found in a subpopulation of cholinergic neurons. Despite this heterogeneity, gene expression patterns in individual cholinergic cells appear to be highly regulated in a cell-specific manner. In fact, membrane receptor transcripts are clustered with their respective intracellular signaling and downstream targets. This novel population of cholinergic neurons may be part of the neural circuitries that detect homeostatic need for food and control the drive to eat.
Division of Endocrinology, Department of Medicine, Albert Einstein College of Medicine, Bronx, United States of America.
Jae Hoon
http://orcid.org/0000-0001-9053-138X
JH
Jeong
Department of Genetics, Albert Einstein College of Medicine, Bronx, United States of America.
Young Jae
YJ
Woo
Division of Endocrinology, Department of Medicine, Albert Einstein College of Medicine, Bronx, United States of America.
Streamson
S
Chua
Division of Endocrinology, Department of Medicine, Albert Einstein College of Medicine, Bronx, United States of America.|Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, United States of America.
Young-Hwan
YH
Jo
D000818:Animals; D001111:Arcuate Nucleus of Hypothalamus / Q000166:cytology* / Q000378:metabolism*; D059329:Cholinergic Neurons / Q000378:metabolism*; D016000:Cluster Analysis; D020869:Gene Expression Profiling / Q000379:methods*; D008810:Mice, Inbred C57BL; D011333:Pro-Opiomelanocortin / Q000378:metabolism; D054411:Receptors, Leptin / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D054411:Receptors, Leptin; D011333:Pro-Opiomelanocortin; D014446:Tyrosine 3-Monooxygenase