This study applied single-nucleus RNA-seq (snRNA-seq) and multiplexed error robust fluorescence in situ hybridization (MERFISH) to resolve neuronal subtypes and map their spatial distribution in the dorsal pons (dPnTg) of mice. The dPnTg is a complex region involved in regulating vital functions, including autonomic nervous system control and integration of sensory, cognitive, and emotional inputs. The study identified over 120 neuronal subtypes, revealing high transcriptional diversity and unique marker genes for many subtypes. The analysis also showed that many neuronal subtypes are transcriptionally similar between humans and mice, enhancing the translational value of the findings. A freely accessible dashboard was developed to provide interactive visual analytics and hardware-accelerated SQL for data exploration. The study's results provide a comprehensive transcriptional atlas of the dPnTg, facilitating future mechanistic investigations into the region's functions.This study applied single-nucleus RNA-seq (snRNA-seq) and multiplexed error robust fluorescence in situ hybridization (MERFISH) to resolve neuronal subtypes and map their spatial distribution in the dorsal pons (dPnTg) of mice. The dPnTg is a complex region involved in regulating vital functions, including autonomic nervous system control and integration of sensory, cognitive, and emotional inputs. The study identified over 120 neuronal subtypes, revealing high transcriptional diversity and unique marker genes for many subtypes. The analysis also showed that many neuronal subtypes are transcriptionally similar between humans and mice, enhancing the translational value of the findings. A freely accessible dashboard was developed to provide interactive visual analytics and hardware-accelerated SQL for data exploration. The study's results provide a comprehensive transcriptional atlas of the dPnTg, facilitating future mechanistic investigations into the region's functions.