Hippocampal Cell Culture and conditions
Rat primary hippocampal neuronal cells were purchased from QBM Cell
Science (Montreal, Canada) and cultured in Neuro basal medium
supplemented with 10% FBS, 2 mM GlutaMAX-1, 1 mM sodium pyruvate, 100
U/ml penicillin G, and 100 μg/ml streptomycin and 2% B-27 supplement at
37°C in a humidified 10% CO2 environment according to
manufacturer’s instructions. Half part of the medium was changed every 3
days. The cells were grown under the above conditions (control
condition) for 7 d. Cultured hippocampal neurons were grown and treated
for 24 hours under specific pharmacologic conditions. Treated
hippocampal cells were divided into experimental groups: (1) Control
group; (2) M1145-treated group (100nM); (3) Y1R agonist-treated group
receiving an NPYY1R agonist
[Leu31,Pro34]NPY (100nM); (4)
GAL+Y1R: Group administered with both substances; (5) GAL+Y1+M871: Group
injected with GAL,
[Leu31,Pro34]NPY and the GALR2
antagonist (M871; 1μM). Cells were grown on poly-D-lysine-coated glass
coverslips and fixed with 4% formaldehyde solution for 20 min followed
by two washes with PBS containing 20 mM glycine to quench the aldehyde
groups.