The localization of the neuropeptide tyrosine (NPY) Y2 receptor (Y2R) protein was studied in mouse dorsal root ganglia (DRGs) and spinal cord, by using a recently developed rabbit anti‐Y2R antibody and a sensitive immunohistochemical method. Y2R‐like immunoreactivity (‐LI) was observed in about 10% of the small/medium‐sized lumbar DRG neurons. Among these, about 44% were calcitonin gene‐related peptide‐immunoreactive, and about 38% bound isolectin B4. In the dorsal horn of the spinal cord, an intense Y2R‐LI was seen in the most superficial layers, mostly restricted to laminae I–II. This immunoreactivity was completely abolished by dorsal rhizotomy. Y2R‐L1 was also detected on the skin, more abundantly in hairy than glabrous skin. Specificity experiments showed complete disappearance of the Y2R‐LI described above after incubation with antibody preadsorbed with the immunogenic peptide. Furthermore, Y2R‐LI was also absent in a Y2R knockout mouse. These results demonstrate that the NPY Y2R is associated mainly with both peptidergic and nonpeptidergic small, presumably nociceptive, neurons projecting to the superficial layers of the dorsal horn. The results also support a role for this receptor and NPY in pain mechanisms. J. Comp. Neurol. 489:328–348, 2005. © 2005 Wiley‐Liss, Inc.