Future studies may benefit from investigating the effect of alcohol under such conditions. Third, our puff application of neurotransmitters produced a local dilution of alcohol during recordings to examine alteration of receptor responses by bath-applied alcohol. Use of a multi-barrel pipette for puff application could limit dilution of local alcohol concentration during receptor activation in such experiments. Fourth, as with any in vitro study, the neural cells we examined were not in their native environment and variation in culture conditions or well-to-well variability may impact patterns of gene expression (Newman and Cooper, 2010). Fifth, although the genetic sequence differences contributing to risk of alcohol dependence in neural cells derived from AD individuals are likely to be maintained in iPS cells, the environmental effect of chronic alcohol exposure in AD donor subjects (e.g. epigenetic, transcriptional or post-translational effects of alcohol on neural cells) are not likely to be preserved during reprogramming. Sixth, the results generated from our pilot data do not identify whether changes in mRNA expression directly lead to changes in NMDA receptor activity. Post-translational effects of alcohol on NMDA receptors have also been reported. Fyn kinase-mediated phosphorylation of tyrosine residues on the long intracellular tail of the NR2B subunit
