Abstract Objective Phenylethanolamine N ‐methyltransferase (PNMT)‐expressing neurons in the nucleus tractus solitarii (NTS) contribute to the regulation of autonomic functions. However, the neural circuits linking these neurons to other brain regions remain unclear. This study aims to investigate the connectivity mechanisms of the PNMT‐expressing neurons in the NTS (NTS PNMT neurons). Methods The methodologies employed in this study included a modified rabies virus‐based retrograde neural tracing technique, conventional viral anterograde tracing, and immunohistochemical staining procedures. Results A total of 43 upstream nuclei projecting to NTS PNMT neurons were identified, spanning several key brain regions including the medulla oblongata, pons, midbrain, cerebellum, diencephalon, and telencephalon. Notably, dense projections to the NTS PNMT neurons were observed from the central amygdaloid nucleus, paraventricular nucleus of the hypothalamus, area postrema, and the gigantocellular reticular nucleus. In contrast, the ventrolateral medulla, lateral parabrachial nucleus, and lateral hypothalamic area were identified as the primary destinations for axon terminals originating from NTS PNMT neurons. Additionally, reciprocal projections were evident among 21 nuclei, primarily situated within the medulla oblongata. Conclusion Our research findings demonstrate that NTS PNMT neurons form extensive connections with numerous nuclei, emphasizing their essential role in the homeostatic regulation of vital autonomic functions.