Figure 6

Gene expression in dispersed hypothalamic neurons isolated from chicks following feeding or fasting for 24 or 48 h and treated with receptor agonists in culture. Following feeding or fasting of chicks for 24 or 48 h, the hypothalamus was collected and neurons were dispersed and cultured in 12-well culture dishes in neurobasal medium for 24 h. Receptor agonists [α- melanocyte stimulating hormone (aMSH), neuropeptide Y (NPY), somatostatin (SRIF), norepinephrine (NE), or O-phospho-L-serine (LSOP, a group III metabotropic receptor agonist] were added to the medium following 24 h of incubation, and neurons were incubated for an additional 24 h. Cells were collected, and RNA was extracted for analysis by qRT-PCR. Gene expression was evaluated for pro-opiomelanocortin (POMC), melanocortin 4 receptor (MC4R), neuropeptide Y (NPY), beta-2-adrenergic receptor (ADRB2), and metabotropic glutamate receptor 8 (GRM8). The first column compares the relative gene expression levels between fed and fasted chicks within each time point (24 h - white bars, 48 h - black bars). Gene expression levels in treated neurons from chicks fed (black bars) or fasted (white bars) for 24 h are represented in the 2nd column, and levels from chicks fed (black bars) or fasted (white bars) for 48 h are in the 3rd column. Asterisks indicate significant differences (P < 0.05) within time points for the control graphs (column 1) or within time and nutritional status (fed or fasted) (columns 2 and 3).