Itch intensity, quality and characteristics differ between body regions. Despite their prevalence, the mechanisms underlying regional itch differences remain poorly understood. Here, we investigate the neural basis of regional differences, focusing on the functional role of neuropeptides and histamine receptors expression in trigeminal ganglia versus dorsal root ganglia innervating afferents to cheek and the neck. Using an interdisciplinary approach, we identified distinct regional differences between the cheeks and necks of mice and humans. Our findings reveal that Substance P modulates itch in the cheek, while histamine receptor 1 (HRH1) is the primary receptor mediating itch but not pain in the cheek. We also discovered regional differences in sensory neuron subpopulations, with increased co-expression of HRH1 and neuropeptides in trigeminal ganglion neurons. Our study provides new insights into the molecular and cellular mechanisms underlying regional itch differences, supporting region-specific treatment strategies for skin and systemic diseases.