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

MB Allison, CM Patterson, MJ Krashes, BB Lowell… - Molecular …, 2015 - Elsevier
MB Allison, CM Patterson, MJ Krashes, BB Lowell, MG Myers Jr, DP Olson
Molecular metabolism, 2015Elsevier
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 …
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, Pdyncre-mediated ablation of Leprflox in Pdyn-expressing neurons (LepRbPdynKO 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