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NPY also acts in the mesolimbic dopaminergic circuitry, where it can increase motivational aspects of feeding behavior through effects on dopamine output in the nucleus accumbens (NAc) and on neurotransmission in the ventral tegmental area (VTA). Endogenous NPY in the NAc originates from local interneurons and afferent projections from the hypothalamic arcuate nucleus (Arc). However, the origin of endogenous NPY in the VTA is unknown. We determined, in normal\u2010weight male Wistar rats, if the source of VTA NPY is local, and\/or whether it is derived from VTA\u2010projecting neurons. Immunocytochemistry, <jats:italic>in situ<\/jats:italic> hybridization and RT\u2010qPCR were utilized, when appropriate in combination with colchicine treatment or 24\u2009hr fasting, to assess NPY\/<jats:italic>Npy<\/jats:italic> expression locally in the VTA. Retrograde tracing using cholera toxin beta (CTB) in the VTA, fluorescent immunocytochemistry and confocal microscopy were used to determine NPY\u2010immunoreactive afferents to the VTA. NPY in the VTA was observed in fibers, but not following colchicine pretreatment. No NPY\u2010 or <jats:italic>Npy<\/jats:italic>\u2010expressing cell bodies were observed in the VTA. Fasting for 24\u2009hr, which increased <jats:italic>Npy<\/jats:italic> expression in the Arc, failed to induce <jats:italic>Npy<\/jats:italic> expression in the VTA. Double\u2010labeling with CTB and NPY was observed in the Arc and in the ventrolateral medulla. Thus, VTA NPY originates from the hypothalamic Arc and the ventrolateral medulla of the brainstem in normal\u2010weight male Wistar rats. These afferent connections link hypothalamic and brainstem processing of physiologic state to VTA\u2010driven motivational behavior.<\/jats:p>","DOI":"10.1002\/cne.24698","type":"journal-article","created":{"date-parts":[[2019,4,6]],"date-time":"2019-04-06T01:33:41Z","timestamp":1554514421000},"page":"2659-2674","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Afferent neuropeptide Y projections to the ventral tegmental area in normal\u2010weight male Wistar rats"],"prefix":"10.1002","volume":"527","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4347-6300","authenticated-orcid":false,"given":"Myrtille C. 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