Cells were plated at a 60,000 cells/cm2 density onto poly D-lysine-coated coverslips after electroporation and cultured to maturity (DIV14-DIV17) before imaging. Neurons were imaged live using an inverted motorized epifluorescence microscope (Olympus,IX81) fitted with a Prime 95B camera. The coverslips were mounted into a Chamlide EC magnetic chamber
(Live cell Instrument, ON, Canada) in Tyrode solution (pH 7.4) containing (in mM):
119 NaCl, 2.5 KCl, 2 CaCl2, 2 MgCl2, 25 HEPES, 30 glucose, 10 μM 6-cyano-7-nitroquinoxaline-2,3-dione 015 (CNQX, TOCRIS bioscience #0190), and 50 μM D,L-2-amino-5-phosphonovaleric acid (AP5, TOCRIS bioscience #0105).NH4Cl perfusions were done with 50mM NH4Cl in substitution of 50 mM NaCl (pH 7.4).