title: TRAP-seq defines markers for novel populations of hypothalamic and brainstem LepRb neurons.
abstract: 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.
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA ; Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Margaret B
authors.initials: MB
authors.lastname: Allison
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA ; Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Christa M
authors.initials: CM
authors.lastname: Patterson
authors.affiliation: Division of Endocrinology, Beth Israel-Deaconess Medical Center, Boston, MA, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Krashes
authors.affiliation: Division of Endocrinology, Beth Israel-Deaconess Medical Center, Boston, MA, USA.
authors.forename: Bradford B
authors.initials: BB
authors.lastname: Lowell
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA ; Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Martin G
authors.initials: MG
authors.lastname: Myers
authors.affiliation: Division of Endocrinology, Department of Pediatrics and Communicable Diseases, University of Michigan, 1000 Wall St, 6321 Brehm Tower, Ann Arbor, MI 48105, USA.
authors.forename: David P
authors.initials: DP
authors.lastname: Olson
keywords: Brainstem; Dynorphin; Hypothalamus; Leptin; Neuropeptides
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