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TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons

OBJECTIVE: Leptin acts via its receptor (LepRb) on multiple subpopulations of LepRb neurons in the brain, each of which controls specific aspects of energy balance. Despite the importance of LepRb-containing neurons, the transcriptome and molecular identity of many LepRb subpopulations remain undefi...

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Autores principales: Allison, Margaret B., Patterson, Christa M., Krashes, Michael J., Lowell, Bradford B., Myers, Martin G., Olson, David P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354921/
https://www.ncbi.nlm.nih.gov/pubmed/25830093
http://dx.doi.org/10.1016/j.molmet.2015.01.012
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author Allison, Margaret B.
Patterson, Christa M.
Krashes, Michael J.
Lowell, Bradford B.
Myers, Martin G.
Olson, David P.
author_facet Allison, Margaret B.
Patterson, Christa M.
Krashes, Michael J.
Lowell, Bradford B.
Myers, Martin G.
Olson, David P.
author_sort Allison, Margaret B.
collection PubMed
description OBJECTIVE: Leptin acts via its receptor (LepRb) on multiple subpopulations of LepRb neurons in the brain, each of which controls specific aspects of energy balance. Despite the importance of LepRb-containing neurons, the transcriptome and molecular identity of many LepRb subpopulations remain undefined due to the difficulty of studying the small fraction of total cells represented by LepRb neurons in heterogeneous brain regions. Here we sought to examine the transcriptome of LepRb neurons directly and identify markers for functionally relevant LepRb subsets. METHODS: We isolated mRNA from mouse hypothalamic and brainstem LepRb cells by Translating Ribosome Affinity Purification (TRAP) and analyzed it by RNA-seq (TRAP-seq). RESULTS: TRAP mRNA from LepRb cells was enriched for markers of peptidergic neurons, while TRAP-depleted mRNA from non-LepRb cells was enriched for markers of glial and immune cells. Genes encoding secreted proteins that were enriched in hypothalamic and brainstem TRAP mRNA revealed subpopulations of LepRb neurons that contained neuropeptide-encoding genes (including prodynorphin, Pdyn) not previously used as functional markers for LepRb neurons. Furthermore, Pdyn(cre)-mediated ablation of Lepr(flox) in Pdyn-expressing neurons (LepRb(Pdyn)KO mice) blunted energy expenditure to promote obesity during high-fat feeding. CONCLUSIONS: TRAP-seq of CNS LepRb neurons defines the LepRb neuron transcriptome and reveals novel markers for previously unrecognized subpopulations of LepRb neurons.
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spelling pubmed-43549212015-03-31 TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons Allison, Margaret B. Patterson, Christa M. Krashes, Michael J. Lowell, Bradford B. Myers, Martin G. Olson, David P. Mol Metab Original Article OBJECTIVE: Leptin acts via its receptor (LepRb) on multiple subpopulations of LepRb neurons in the brain, each of which controls specific aspects of energy balance. Despite the importance of LepRb-containing neurons, the transcriptome and molecular identity of many LepRb subpopulations remain undefined due to the difficulty of studying the small fraction of total cells represented by LepRb neurons in heterogeneous brain regions. Here we sought to examine the transcriptome of LepRb neurons directly and identify markers for functionally relevant LepRb subsets. METHODS: We isolated mRNA from mouse hypothalamic and brainstem LepRb cells by Translating Ribosome Affinity Purification (TRAP) and analyzed it by RNA-seq (TRAP-seq). RESULTS: TRAP mRNA from LepRb cells was enriched for markers of peptidergic neurons, while TRAP-depleted mRNA from non-LepRb cells was enriched for markers of glial and immune cells. Genes encoding secreted proteins that were enriched in hypothalamic and brainstem TRAP mRNA revealed subpopulations of LepRb neurons that contained neuropeptide-encoding genes (including prodynorphin, Pdyn) not previously used as functional markers for LepRb neurons. Furthermore, Pdyn(cre)-mediated ablation of Lepr(flox) in Pdyn-expressing neurons (LepRb(Pdyn)KO mice) blunted energy expenditure to promote obesity during high-fat feeding. CONCLUSIONS: TRAP-seq of CNS LepRb neurons defines the LepRb neuron transcriptome and reveals novel markers for previously unrecognized subpopulations of LepRb neurons. Elsevier 2015-02-07 /pmc/articles/PMC4354921/ /pubmed/25830093 http://dx.doi.org/10.1016/j.molmet.2015.01.012 Text en © 2015 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Allison, Margaret B.
Patterson, Christa M.
Krashes, Michael J.
Lowell, Bradford B.
Myers, Martin G.
Olson, David P.
TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons
title TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons
title_full TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons
title_fullStr TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons
title_full_unstemmed TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons
title_short TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons
title_sort trap-seq defines markers for novel populations of hypothalamic and brainstem leprb neurons
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354921/
https://www.ncbi.nlm.nih.gov/pubmed/25830093
http://dx.doi.org/10.1016/j.molmet.2015.01.012
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