An in vitro model of human neocortical development using pluripotent stem cells: cocaine-induced cytoarchitectural alterations.
Bottom Line: This model was used to examine the effects of cocaine during neocorticogenesis.These cocaine-induced changes were inhibited by the cytochrome P450 inhibitor cimetidine.This in vitro model enables mechanistic studies of neocorticogenesis, and can be used to examine the mechanisms through which cocaine alters the development of the human neocortex.
Affiliation: Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS), Baltimore, MD 21244, USA.Show MeSH
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Mentions: To study how cocaine affects neocorticogenesis and the mechanisms involved in cytochrome-P450-mediated N-oxidative metabolism of cocaine in humans, we first examined whether cocaine induces ROS formation in NE cells derived from hPSCs on day 19. As shown in Fig. 4A, a pharmacologically relevant dose of cocaine, 3 μM (Kourrich et al., 2013), caused a significant increase in ROS 30 minutes after cocaine treatment. Pretreatment of NE cells, 30 minutes prior to cocaine exposure, with the cytochrome P450 inhibitor cimetidine, which blocks the N-oxidative metabolism of cocaine, inhibited cocaine-induced ROS formation (Fig. 4A). We previously reported that cocaine concentrations in the fetal rat brain decrease rapidly less than 1 hour after cocaine injection (Lee et al., 2008). Therefore, in order to create a physiologically meaningful drug treatment schedule in vitro, hPSC-derived neocortical cells were treated with 3 μM cocaine for 1 hour at days 20, 22 and 24, the beginning of neocortical neurogenesis, during which the NE cells are actively cycling.
Affiliation: Cellular Neurobiology Research Branch, Intramural Research Program (IRP), National Institute on Drug Abuse, National Institutes of Health (NIH), Department of Health and Human Services (DHHS), Baltimore, MD 21244, USA.