Exposure to amino acid neurotransmitters causes cell death in early GFP+ cultures.
Cultures containing Pitx3-eGFP+ cells were incubated with the membrane impermeable dye TOPRO-3 and exposed to GABA (30 µM), Glut (30 µM), or a combination of the two. (A) shows the effects of GABA exposure on cells at day 15 and day 23. The panels show, from top to bottom, eGFP and TOPRO-3 at time = 0, after 3 hours of constant incubation, and overlays of regions showing increased TOPRO-3 reactivity. After 3 hours of amino acid incubation there were a significantly higher number of red overlays in non-vehicle control treated cells from day 15 (One way ANOVA with post-hoc Dunnett's test, P<0.05, n = 5) (B) than were present in cultures from day 23 (C).