A spatially-resolved transcriptional atlas of the murine dorsal pons at single-cell resolution was developed using single-nucleus RNA sequencing (snRNA-seq) and multiplexed error-robust fluorescence in situ hybridization (MERFISH). The study identified over 120 neuronal subtypes in the dorsal pons (dPnTg), revealing high transcriptional diversity and defining unique marker genes for many subtypes. The analysis also demonstrated significant transcriptional similarity between human and mouse neurons, enhancing the translational value of the study. A freely accessible dashboard was developed to enable data exploration and analysis. The dPnTg is a complex region of the brainstem involved in regulating vital functions such as respiration, arousal, sleep-wake regulation, pain, reward processing, movement, memory, feeding, micturition, aversive behaviors, thermoregulation, and cardiovascular regulation. The study focused on the dPnTg, which is part of the pontine tegmentum and plays a critical role in the autonomic nervous system. The research identified 63 clusters of cells, including 12 major cell types, with neurons accounting for ~40% of all nuclei. The study also identified 11 major glial/non-neuronal cell types. The analysis of the dPnTg revealed distinct neuronal subtypes, including excitatory and inhibitory neurons, with unique marker genes. The study also provided a detailed atlas of the parabrachial nucleus (PB), the Kölliker Fuse (KF), the medial pontine tegmental nucleus (MTN), the locus coeruleus (LC), and the Bar nucleus. The study identified 44 clusters of cells in the dPnTg, with neurons accounting for 50% of all cells. The study also identified 27 clusters in the LDTg/LDTgV region, with 17 being GABAergic, 8 glutamatergic, and 2 cholinergic. The study demonstrated that many neuronal subtypes are transcriptionally similar between humans and mice, enhancing the translational value of the study. The study also identified a high degree of transcriptional similarity between mouse and human neuronal subtypes, indicating evolutionary conservation of function in this brain region. The study provided a detailed atlas of the dPnTg, including the KF, PB, MTN, LC, and Bar, and identified unique marker genes for each subregion. The study also identified 36 clusters in the dPnTg, with distinct marker genes for each cluster. The study demonstrated that the dPnTg is a complex region with a high degree of transcriptional diversity and that many neuronal subtypes are transcriptionally similar between humans and mice. The study also provided a detailed atlas of the dPnTg, including the KF, PB, MTN, LCA spatially-resolved transcriptional atlas of the murine dorsal pons at single-cell resolution was developed using single-nucleus RNA sequencing (snRNA-seq) and multiplexed error-robust fluorescence in situ hybridization (MERFISH). The study identified over 120 neuronal subtypes in the dorsal pons (dPnTg), revealing high transcriptional diversity and defining unique marker genes for many subtypes. The analysis also demonstrated significant transcriptional similarity between human and mouse neurons, enhancing the translational value of the study. A freely accessible dashboard was developed to enable data exploration and analysis. The dPnTg is a complex region of the brainstem involved in regulating vital functions such as respiration, arousal, sleep-wake regulation, pain, reward processing, movement, memory, feeding, micturition, aversive behaviors, thermoregulation, and cardiovascular regulation. The study focused on the dPnTg, which is part of the pontine tegmentum and plays a critical role in the autonomic nervous system. The research identified 63 clusters of cells, including 12 major cell types, with neurons accounting for ~40% of all nuclei. The study also identified 11 major glial/non-neuronal cell types. The analysis of the dPnTg revealed distinct neuronal subtypes, including excitatory and inhibitory neurons, with unique marker genes. The study also provided a detailed atlas of the parabrachial nucleus (PB), the Kölliker Fuse (KF), the medial pontine tegmental nucleus (MTN), the locus coeruleus (LC), and the Bar nucleus. The study identified 44 clusters of cells in the dPnTg, with neurons accounting for 50% of all cells. The study also identified 27 clusters in the LDTg/LDTgV region, with 17 being GABAergic, 8 glutamatergic, and 2 cholinergic. The study demonstrated that many neuronal subtypes are transcriptionally similar between humans and mice, enhancing the translational value of the study. The study also identified a high degree of transcriptional similarity between mouse and human neuronal subtypes, indicating evolutionary conservation of function in this brain region. The study provided a detailed atlas of the dPnTg, including the KF, PB, MTN, LC, and Bar, and identified unique marker genes for each subregion. The study also identified 36 clusters in the dPnTg, with distinct marker genes for each cluster. The study demonstrated that the dPnTg is a complex region with a high degree of transcriptional diversity and that many neuronal subtypes are transcriptionally similar between humans and mice. The study also provided a detailed atlas of the dPnTg, including the KF, PB, MTN, LC