Neurosteroid Binding Sites on the GABAA Receptor Complex as Novel Targets for Therapeutics to Reduce Alcohol Abuse and Dependence
Figure 1
The amount of DHEA (ng/g) present in the hypothalamus, hippocampus, and frontal cortex of rats over a six-hour time period after a single acute intraperitoneal injection. Adult male Long-Evans rats received either 56 mg/kg of DHEA () or an equal volume of cyclodextrin vehicle (). A DHEA-treated subject was sacrificed with a vehicle-treated control at 15, 30, 60, 120, and 180 minutes after injection, while the final DHEA-treated subject was sacrificed 360 minutes after injection. Brains were collected, flash frozen, and later dissected using the Glowinski technique [75]. Steroids were extracted from the hypothalamus, hippocampus, and frontal cortex of each subject using the solid-phase extraction method described and validated by Newman et al. [74]. Briefly, tissue from each region was prepared in an aqueous matrix and steroids were extracted from each sample using a C18 column primed with ethanol and equilibrated with water. Each sample was eluted, dried, and resuspended in deionized water, and DHEA levels were determined using ELISA.
