22Corticotropin-releasing hormone (CRH) is an important neuromodulator with wide 23 distribution in the brain. Here, we screened the CRH-IRES-Cre;Ai32 mouse line to reveal 24 the morphologies of individual CRH neurons throughout the mouse brain by using 25 fluorescence micro-optical sectioning tomography (fMOST) system. Diverse dendritic 26 morphologies and projection fibers were found in various brain regions. Reconstructions 27 showed hypothalamic CRH neurons had the smallest somatic volumes and simplest 28 dendritic branches, and CRH neurons in several regions shared a bipolar morphology. 29 Further investigations in the medial prefrontal cortex unveiled somatic depth-dependent 30 morphologies that exhibited three types of connections and CRH neurons in the anterior 31 parvicellular area of hypothalamus had fewer and smaller Herring bodies whereas in the 32 periventricular area had more and larger Herring bodies that were present within fibers 33 projecting to the third ventricle. Our findings provide the most comprehensive intact 34 morphologies of CRH neurons throughout the mouse brain that is currently available. 35 36 42 neurons are also broadly distributed in other brain regions, including the inferior olivary 43 nucleus, Barrington's nucleus, pontine tegmentum, cerebral cortex, hippocampus, and 44 central amygdala. Depending on the region-specific somatic locations, CRH neurons 45 participate in various functional activities, such as learning memory, synaptic plasticity, 46 food intake and drug addiction, as well as anxiety-like and depression-like behaviors 47 Charlton et al., 1987;Cummings, 1989;Delville et al., 1992;Foote and Cha, 1988; 51 Merchenthaler, 1984;Shu et al., 2015). However, data from these studies have mainly been 52 acquired from histological imaging and through manual reconstruction and counting of 53 labeled neurons, which is time-consuming, limits further systematic analysis, and can 54 introduce biases and/or artifacts. Recently, genetically modified mouse models have been 55 developed to identify the whole-brain distributions of CRH neurons (Itoi et al., 2014; Kono 56 et al., 2017; Peng et al., 2017; Wamsteeker Cusulin et al., 2013), which has significantly 57 advanced our understanding of the morphological features of CRH neurons in the rodent 58 brain (De Francesco et al., 2015; Garcia et al., 2016; Huang et al., 2013; Nguyen et al., 59 2016). Advances in whole-brain optical imaging techniques, such as fluorescence micro-60 optical sectioning tomography (fMOST) (Gong et al., 2016; Gong et al., 2013; Ragan et 61 al., 2012; Zheng et al., 2013), have made it feasible to further quantify cellular distributions 62 and to morphologically reconstruct cells at the whole-brain level. The precision of imaging 63 via fMOST can reveal complex fiber orientations, and can even distinguish individual 64 dendrites. Such quantitative three-dimensional (3D) neuronal morphologies obtained at a 65 brain-wide scale can provide highly accurate arborization details and comprehensive 66 mapping of...