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. Author manuscript; available in PMC: 2012 Mar 1.

Published in final edited form as: Pharmacol Ther. 2010 Oct 21;129(3):241–259. doi: 10.1016/j.pharmthera.2010.09.010

Figure 1A. Warm sensitive neurons in the Preoptic area that were subject of single cell transcriptomics study by chipping and sequencing. Thermoregulatory network is depicted showing the central integrative role of temperature sensitive GABAergic projection neurons (highlighted in red) in activation of thermogenesis in Brown Adipose Tissue, respiratory rate and cardiac output. It depicts data from multiple retrograde tracing studies that show that the inhibitory warm sensitive neurons project to DMH and or rRPA., and that the skin temperature and deep body temperature is affecting the activity of these neurons through Glutamatergic afferents while the local temperature is belived to exert its effect through circulation communicated heat affects. POA: Preoptic Area, DMH: Dorsomedial Hypothalamus, rRPA: rostral Raphe Pallidus, DRG: Dorsal Root Ganglion, BAT: Brown Adipose Tissue, LPB: Lateral parabrachial nucleus. Adapted from Nakamura, K. and S. F. Morrison (2008) and modified with our data.

Figure 1B. Flow chart of experimental design for single cell transcriptomics-based identification and in vitro and in vivo validation of the functional secretome and receptor repertoire of individual warm sensitive neurons. The numbers in red show the number of transcripts identified in individual neuron cDNA libraries by chipping using the Affymetrix mouse microarray, in the warm sensitive neurons, the number of cDNAs confirmed by NextGen sequencing that produces reads of 48–100 nucleotides, and accepting reads that are fully matching, in the functional validation experiments involved agonists that were tested electrophysiologically on spontaneously active warm sensitive neurons in slice (at least on ten to fifteen such neurons), 31of these agonist for recptors whose cDNA we identified, were found able to affect the behavior of the cells, i.e., have functional receptors expressed on warm sensitive cells, and the remaining ones may not havefunctional receptors expressed or the receptors do not affect the tested electrophysiological properties, or the number of cells tested was too low to find a cell with a functional receptor expressed, 21 recptor agonists were injected into the POA as cDNA encoding their receptor, or channel was found, 18 of these produced hypo- or hyperthermic response at the 0.01 to 10 nanomoles dose.

Figure 1A. Warm sensitive neurons in the Preoptic area that were subject of single cell transcriptomics study by chipping and sequencing. Thermoregulatory network is depicted showing the central integrative role of temperature sensitive GABAergic projection neurons (highlighted in red) in activation of thermogenesis in Brown Adipose Tissue, respiratory rate and cardiac output. It depicts data from multiple retrograde tracing studies that show that the inhibitory warm sensitive neurons project to DMH and or rRPA., and that the skin temperature and deep body temperature is affecting the activity of these neurons through Glutamatergic afferents while the local temperature is belived to exert its effect through circulation communicated heat affects. POA: Preoptic Area, DMH: Dorsomedial Hypothalamus, rRPA: rostral Raphe Pallidus, DRG: Dorsal Root Ganglion, BAT: Brown Adipose Tissue, LPB: Lateral parabrachial nucleus. Adapted from Nakamura, K. and S. F. Morrison (2008) and modified with our data.

Figure 1B. Flow chart of experimental design for single cell transcriptomics-based identification and in vitro and in vivo validation of the functional secretome and receptor repertoire of individual warm sensitive neurons. The numbers in red show the number of transcripts identified in individual neuron cDNA libraries by chipping using the Affymetrix mouse microarray, in the warm sensitive neurons, the number of cDNAs confirmed by NextGen sequencing that produces reads of 48–100 nucleotides, and accepting reads that are fully matching, in the functional validation experiments involved agonists that were tested electrophysiologically on spontaneously active warm sensitive neurons in slice (at least on ten to fifteen such neurons), 31of these agonist for recptors whose cDNA we identified, were found able to affect the behavior of the cells, i.e., have functional receptors expressed on warm sensitive cells, and the remaining ones may not havefunctional receptors expressed or the receptors do not affect the tested electrophysiological properties, or the number of cells tested was too low to find a cell with a functional receptor expressed, 21 recptor agonists were injected into the POA as cDNA encoding their receptor, or channel was found, 18 of these produced hypo- or hyperthermic response at the 0.01 to 10 nanomoles dose.