Figure 2.

LS neurons with TMEM16B expression project to ventral LS and hypothalamic nuclei. TMEM16B-expressing LS neurons project to LPO and LHA. A, Modified Allen Brain Reference Atlas images with locations of LPO (left) and LHA (right) delineated by black border. B, Modified Allen Brain ISH images for TMEM16B mRNA (gene name: Ano2; experiment #71587834) (right). Black border indicates absence of TMEM16B mRNA expression in LPO (left) and LHA (right), suggesting that mCherry expression is from LS nerve fibers projecting to these hypothalamic regions. Arrows point to the specific brain region. C, mCherry fluorescence images in coronal section of TMEM16B KO mouse brain. Large white border indicates location of mCherry axon fibers in LPO (left) and LHA (right). Small white borders indicate the caudal LS (LSc), which is part of the dorsal regions of the LS, and the LSv. Arrows point to the specific brain region. D, 63× magnification image of the LSv with DAPI and mCherry labeling. mCherry labels the nerve fibers in the LSv but not the cell membranes surrounding DAPI-labeled cell bodies. E, 63× magnification image of the LSc with DAPI and mCherry labeling. mCherry puncta localized to the cell membranes surrounding DAPI-labeled cell bodies (see arrowheads). Scale bars: 63× images, 20 μm; all coronal section images, 1000 μm.