Extended Data Fig. 4: Parabrachial neurons innervate tanycytes.
From: A brainstem–hypothalamus neuronal circuit reduces feeding upon heat exposure
a. Confocal image of GFP+ neurons within the arcuate nucleus (ARC; in cyan) transduced with an AAVrg-CAG-GFP vector and positioned ventrolateral to the 3rd ventricle (3V). Scale bar = 100 µm. ‘1’. Panoramic image from Fig. 2e, showing GFP+ neurons (in cyan) in the locus coeruleus (LC) and parabrachial nucleus (PBN) retrogradely labelled with an AAVrg-CAG-GFP construct. Scale bar = 100 µm. b. Panoramic confocal image of a GFP+ neuron (in cyan). Open rectangle denotes the position of ‘1’ in the PBN trans-synaptically labelled by rAAV8-EF1a-mCherry-IRES-WGA-Cre in TaumGFP-loxP mice. Scale bar = 50 µm (b), 8 µm (1). c. Tiled image survey of Fig. 2f showing GFP+ neurons (in cyan, arrows) in the PBN trans-synaptically labelled by a rAAV8-EF1a-mCherry-IRES-WGA-Cre virus in Ai14 mice. Scale bar = 50 µm. Abbreviations: 4V, 4th ventricle; Aq, aqueduct; scp, superior cerebellar peduncle. d. Number of cFOS+ mCherry-transduced neurons in the PBN. Data in bar graphs represent means ± s.e.m. for saline (grey, n = 5) and CNO (5 mg/kg in red, n = 6) groups. Circles correspond to individual data points; t = 4.094; p = 0.003; Student’s t-test. e. Confocal images of the 3rd ventricle (3V) showing periventricular PBN efferents (mCherry, in magenta) with vimentin+ tanycytes in green. Left and right images are from coordinates −1.94 mm and −2.30 mm (relative to bregma), respectively. Scale bars = 70 µm.
