Sensory neurons innervating the heart transmit chemical and mechanical cues to the central nervous system via the dorsal rootganglion (DRG) and nodose ganglion (NG). Despite their importance in cardiac pain and cardiovascular reflexes, the molecularand functional properties of heart-specific sensory (HS) neurons remain elusive. Here, DRGHS and NGHS neurons innervatingthe heart were FACS purified using a retrograde labeling strategy with Di-8-ANEPPQ, then characterized molecularly by bulkRNA sequencing or evaluated functionally in cocultures with neonatal cardiomyocytes. DRGHS and NGHS neurons formed func-tional connections with neonatal cardiomyocytes as demonstrated by immunolabeling, electron microscopy, and optogeneticmanipulation. Functionally coupled DRGHS neurons exhibited enhanced spontaneous and evoked Ca2+ activity in responseto optogenetic and chemical stimulation, indicating dynamic neuro-cardiac communication. Global RNA sequencing analysisrevealed that DRGHS and NGHS neurons exhibited distinct transcriptomic profiles, including enrichment of transcripts encodingion channels and G protein-coupled receptors, compared with their respective total populations. In DRGHS tissue, these includedScn10a, P2xr2, and Mrgprd, whereas NGHS neurons preferentially expressed P2xr2, Ptgdr, and Cckar, collectively supporting acombination of molecular signatures including nociceptors. Collectively, these findings define molecularly distinct sensory path-ways connecting the heart and the sensory nervous system, providing mechanistic insights and highlighting potential targets formodulation of cardiovascular reflexes and hemostasis.