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Leptin Receptor Long-form Splice-variant Protein Expression in Neuron Cell Bodies of the Brain and Co-localization with Neuropeptide Y mRNA in the Arcuate Nucleus
Abstract
Reduced leptin (Ob protein) signaling is proposed to be a stimulus for the activation of neuropeptide Y (NPY) gene activity and increased expression of mRNA for the long form of the leptin receptor (Ob-Rb) in the hypothalamic arcuate nucleus. To determine if Ob-Rb protein is expressed in arcuate nucleus NPY neurons, we developed an affinity-purified polyclonal antibody against amino acids 956-1102 of human Ob-Rb. This antibody specifically recognizes the cytoplasmic tail of Ob-Rb and does not react with shorter leptin-receptor variants. Western immunoblots of Ob-Rb-transfected COS cells showed a single 150-kD band, and immunofluorescence revealed intense perinuclear staining in the cytoplasm. A 150-kD band was also present in Western immunoblots of hypothalamus. Immunocytochemical staining of brain slices revealed immunoreactive Ob-Rb protein concentrated in many neuronal cell bodies in the same regions of the forebrain that also express Ob-Rb mRNA. In the hypothalamus, Ob-Rb-positive cell bodies were abundant in the arcuate nucleus and ventromedial nucleus, with lesser numbers in the dorsomedial nucleus and paraventricular nucleus. Immunostaining was also detected in cell bodies of pyramidal cell neurons of the pyriform cortex and cerebral cortex, in neurons of the thalamus, and on the surface of ependymal cells lining the third ventricle. The choroid plexus, which expresses the short Ob-Ra form, was negative. Combined immunocytochemistry for Ob-Rb protein and fluorescence in situ hybridization for NPY mRNA identified arcuate nucleus neurons containing both NPY mRNA and Ob-Rb protein. The present finding of Ob-Rb protein in neurons that express NPY mRNA supports the hypothesis that arcuate nucleus NPY neurons are direct targets of leptin and play an important role in regulation of food intake and body weight.
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“where they act on their respective receptors expressed in key brain areas that control energy balance, glucose metabolism and autonomic function (10, 11, 53).”
From Leptin and the Central Nervous System Control of Glucose Metabolism (2011) · cites this paper as [10]
In the citing paper
“leprb mRNA and protein have both been detected in several hypothalamic areas involved in the control of energy balance and autonomic function, including the ARC, ventromedial nucleus (VMH) and dorsomedial nucleus (DMN), with lesser amounts in the paraventricular nucleus (PVN) and the lateral hypothalamic area (LHA) (10, 53).”
From Leptin and the Central Nervous System Control of Glucose Metabolism (2011) · cites this paper as [10]
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Metadata
- Authors
- Baskin, D. G., Schwartz, M. W., Seeley, R. J., Woods, S. C., Porte, D., Breininger, J. F., Jonak, Z., Schaefer, J., Krouse, M., Burghardt, C., Campfield, L. A., Burn, P., & Kochan, J. P.
- Affiliations
- University of Washington (US)
- Department of Medicine, University of Washington, Seattle, Washington
- VA Puget Sound Health Care System (US)
- Department of Metabolic Diseases, Hoffmann-La Roche, Inc., Nutley, New Jersey
- Corresponding author
- Baskin, Denis G.
- Venue
- Journal of Histochemistry & Cytochemistry
- Publisher
- SAGE Publications
- Published
- 1999-03
- Volume
- 47
- Issue
- 3
- Pages
- 353-362
- ISSN
- 0022-1554 (ISSN-L)
- 1551-5044
- OpenAlex
- W2123497298
- Cited by
- 200
- References
- 53
- Citations by year
- 2023: 2 · 2022: 3 · 2021: 1 · 2020: 3 · 2019: 10 · 2018: 2 · 2017: 3 · 2016: 3 · 2015: 3 · 2014: 7 · 2013: 11 · 2012: 9
- FWCI
- 7.05
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Histochemistry & Cytochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- U.S. Department of Veterans Affairs
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases (DK 12829, DK17844, DK-17047, R01DK012829, P30DK017047)
- Topics
- Life Sciences › Neuroscience › Endocrine and Autonomic Systems › Regulation of Appetite and Obesity
- Health Sciences › Medicine › Reproductive Medicine › Hypothalamic control of reproductive hormones
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
- Keywords
- leptin receptor, Ob-Rb, neuropeptide Y, arcuate nucleus, hypothalamus, neuronal expression, protein localization, cytoplasmic tail, food intake regulation, body weight regulation, leptin signaling, ventromedial hypothalamus, paraventricular nucleus, piriform cortex, ependymal cells, choroid plexus, Ob-Ra
- MeSH terms
- Animals, Antibodies, Arcuate Nucleus of Hypothalamus, Carrier Proteins, Hypothalamus, Immunohistochemistry, Liver, Male, Neurons, Neuropeptide Y, Receptors, Cell Surface, RNA, Messenger, Blotting, Western, Rats, Wistar, Alternative Splicing, In Situ Hybridization, COS Cells, Rats, Receptors, Leptin
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