Project description:A 2-mg frozen section of hypothalamus from a single mouse was homogenized in 200 µL of denaturing solution(7 M urea, 2 M thiourea, 20 mM dithiothreitol) containing cOmplete Mini Protease Inhibitor Cocktail (Roche). Peptide extracts equivalent to 135 µg of tissue were analyzed by LC-MS/MS.
Project description:To uncover the cellular architecture of the mouse ventromedial hypothalamus (VMH), we used single nucleus RNA-seq (snRNA-seq) from wild-type mice.
Project description:Neurons in the hypothalamic arcuate nucleus (ARC) are fundamental regulators of energy homeostasis, growth and reproduction. Spatially overlapping subpopulations of ARC neurons produce specific neuropeptides including melanocortins, opioids, neuropeptide Y, growth hormone releasing hormone and kisspeptin. However, the developmental pathways controlling ARC neuron specification, differentiation and maturation remain poorly understood relative to more structurally ordered brain regions. Here, we analyzed the transcriptomes of individual mouse ARC neurons sorted for expression of a Proopiomelanocortin-Td-dsRed transgene at four embryonic (E11.5, E13.5, E15.5 and E17.5) and two postnatal (P5 and P12) ages. There was an unanticipated complexity of neurochemical subpopulations in the developing ARC characterized by low, medium or high levels of Pomc transcripts and distinct temporal patterns of transcription factor expression. Moreover, sizeable fractions of Pomc-positive neural progenitors differentiated into non-POMC neurons by P12. We conclude that ARC neuronal development is highly dynamic with plasticity that may be susceptible to environmental programming.