Each brain was removed from the skull and placed in a chilled rat brain matrix (ASI Instruments, Houston, Texas). A coronal slice containing the region of interest was removed from the matrix and placed on a chilled petri dish. Dissection was carried out using razor blades and forceps under a dissecting microscope. The brain regions of interest were identified according to The Rat Brain in Stereotaxic Coordinates [28]. The lateral hypothalamus (LH), medial portion of the hypothalamus (MH, include the arcuate nucleus, dorsomedial hypothalamus, and paraventricular nucleus), caudate-putamen (CPu), central amygdal (CeA) and medial/basolateral portions of the amygdala (Me/BLA), ventral tegmental area (VTA), and anterior pituitary (AP) were dissected on ice, homogenized in guanidinium thiocyanate buffer and extracted with acidic phenol and chloroform. After the final ethanol precipitation step, each extract was re-suspended in DEPC-treated H2O and stored at −80°C.
