Neuropeptide Y (NPY)‐expressing neurones in the arcuate nucleus densely innervate many hypothalamic nuclei. To determine the neurochemical phenotype of target neurones for NPY, we studied the immunohistochemical localization of the NPY Y1 receptor (Y1R) in discrete subpopulations of neurones in the rat hypothalamus. Among several tested populations, including hypocretin/orexin‐, melanin‐concentrating hormone (MCH)‐ and nitric oxide synthase (NOS)‐positive neurones, only the latter were found to coexpress the Y1R. Numerous Y1R/NOS‐positive neurones were found as a densely packaged group of cells located ventrolateral to the ventromedial nucleus, forming a band ascending towards the fornix. Lower numbers of Y1R/NOS‐positive neurones were found in the perifornical area and in the peri‐ and paraventricular nuclei. Expression of the Y1R gene was found in the same locations in the mouse by colocalizing β‐galactosidase, a Y1R gene reporter, with NOS in a Y1R knockout mouse. To explore possible downstream targets of NO in the rat hypothalamus, the NO‐regulated molecule cGMP was analysed immunohistochemically after incubation of brain slices with sodium nitroprusside, an NO donor. We observed several cGMP‐positive cell bodies in the arcuate nucleus, cGMP‐positive blood vessels and a cGMP‐positive network of thin fibres, some of which colocalized with choline acetyltransferase.