Peptide delivered by nasalspray can reduce seizure activity, protect neurons in Alzheimer's, epilepsy MCG_AUG JCI_Insight
). The A1R 3i loop and CT peptides used for blocking A1R/neurabin interaction were synthesized at Peptide 2.0 and dissolved in water with the stock concentration at 5 μg/μL. The peptide concentration in the final pull-down assay was 0.1 μg/μL.All cell lines were obtained from American Type Culture Collection . CosM6 cells were cultured in DMEM supplemented with 10% FBS , 100 U/mL penicillin, and 10 μg/mL streptomycin.
In brief, hippocampal neurons were cultured in 24-well plates for 13–14 days in vitro and treated with vehicle; 100 μM glutamate alone; 100 μM glutamate plus 10 μM R-PIA; and 100 μM glutamate, 10 μM R-PIA, and 30 μM LY294002 for 30 minutes at 37°C. Neurons were then washed twice and incubated with regular growth medium at 37°C for an additional 24 hours. Neurons were stained with 4 μM calcein-AM and 4 μM EthD-1 in DPBS and mounted for imaging under a fluorescence microscope.