Identification of neuronal input to the arcuate nucleus (ARH) activated during lactation: implications in the activation of neuropeptide Y neurons.

Activation of the neuropeptide Y (NPY) neuronal system in the arcuate nucleus of the hypothalamus (ARH) during lactation in the rat is likely due to the neural impulses arising from the suckling stimulus. However, the afferent neuronal input to the ARH that is activated during lactation and is responsible for activation of NPY neurons is currently unknown. Previously, using cFos as a marker for neuronal activation, we identified several brain areas in the lactating animals that were activated by the suckling stimulus. Thus, the objective of the present study was to determine if these activated areas observed in the lactating animals project directly into the ARH. The retrograde tracer, fluorogold (FG), was injected into the ARH on day 4 postpartum. Chronically suckled rats were then deprived of their eight-pup litters on day 9 postpartum, and 48 h later, the pups were returned to the females to reinitiate the suckling stimulus for 90 min to induce cFos expression. The animals were then perfused and the brains were subjected to double-label immunohistochemistry to visualize both FG- and cFos-positive cells. Substantial FG/cFos double-labeled cells were found in forebrain regions, including the medial preoptic area, periventricular preoptic area, bed nucleus of the stria terminalis, and the medial amygdala, and in brainstem regions including the lateral parabrachial nucleus, peripeduncular area and ventrolateral medulla. The results of the present study demonstrate that specific areas in the brain are activated during lactation and send direct projections to the ARH. Thus, these areas are potentially important candidates for mediating the activation of the NPY neuronal system in the ARH during lactation.