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Previous transsynaptic tracing studies only mapped orofacial premotor circuits in neonatal mice, but the adult circuits remain unknown as a consequence of technical difficulties. Here, we developed a three-step monosynaptic transsynaptic tracing strategy to identify premotor neurons controlling vibrissa, tongue protrusion, and jaw-closing muscles in the adult mouse. We registered these different groups of premotor neurons onto the Allen mouse brain common coordinate framework (CCF) and consequently generated a combined 3D orofacial premotor atlas, revealing unique spatial organizations of distinct premotor circuits. We further uncovered premotor neurons that simultaneously innervate multiple motor nuclei and, consequently, are likely to coordinate different muscles involved in the same orofacial motor actions. Our method for tracing adult premotor circuits and registering to Allen CCF is generally applicable and should facilitate the investigations of motor controls of diverse behaviors.<\/jats:p>","DOI":"10.7554\/elife.67291","type":"journal-article","created":{"date-parts":[[2021,4,27]],"date-time":"2021-04-27T08:20:08Z","timestamp":1619511608000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.67291","source":"Crossref","is-referenced-by-count":54,"title":["Constructing an adult orofacial premotor atlas in Allen mouse CCF"],"prefix":"10.7554","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0976-7684","authenticated-orcid":true,"given":"Jun","family":"Takatoh","sequence":"first","affiliation":[{"name":"Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology","place":["Cambridge, United States"]},{"name":"Department of Neurobiology, Duke University","place":["Durham, United 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