GABAergic neurosteroids mediate the effects of ethanol on long-term potentiation in rat hippocampal slices.

We previously found that ethanol has complex effects on hippocampal synaptic plasticity, inhibiting long-term potentiation (LTP) and long-term depression by different mechanisms. The block of long-term depression appears to be mediated by effects on N-methyl-d-aspartate receptors, whereas the block of LTP involves augmented inhibition via gamma-aminobutyric acid-A receptors (GABA(A)Rs). To pursue factors contributing to effects on LTP, we examined the ability of various concentrations of ethanol to block LTP in the CA1 region of rat hippocampal slices. Complete LTP block required 60 mm ethanol. LTP block was enhanced at lower ethanol concentrations in the presence of (3alpha5alpha)-3-hydroxypregnan-20-one, a GABA(A)R-potentiating neurosteroid, suggesting that neurosteroids may be important contributors to the effects of ethanol on LTP. Consistent with this, we found that block of LTP by 60 mm ethanol was overcome by coadministration of a cyclodextrin that binds and removes lipophilic neurosteroids. More specifically, treatment of slices with finasteride, an agent that inhibits the synthesis of 5alpha-reduced neurosteroids, or with an agent that inhibits the effects of 5alpha-reduced neurosteroids on GABA(A)Rs overcame the effects of 60 mm ethanol on LTP. Taken together, these results indicate that acute production of GABA(A)R-enhancing neurosteroids plays a key role in mediating the effects of ethanol on LTP.

Effects of neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one on ethanol-mediated paired-pulse depression of population spikes in the CA1 region of rat hippocampal slices.

While it is known that ethanol augments GABA-A receptor mediated inhibition in the central nervous system (CNS), demonstrating direct effects of ethanol on GABA transmission has been difficult in brain slices, suggesting that these preparations may lack factors that are required for ethanol's actions. Recent studies indicate that the GABA-enhancing neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha5alphaP) mediates at least some effects of ethanol in the CNS. In the CA1 region of rat hippocampal slices, we found that 60mM ethanol failed to alter paired pulse depression (PDD) of population spikes (PSs) when paired stimuli were delivered to the Schaffer collateral pathway at an interval of 21ms. Following 2-h preincubation of slices with 100nM 3alpha5alphaP, however, ethanol augmented PS PPD. This effect was not observed in the presence of picrotoxin, a GABA-A receptor antagonist, or ADVASEP-7, a beta-cyclodextrin that binds 3alpha5alphaP. These results indicate that 3alpha5alphaP modulates the inhibitory effects of ethanol on hippocampal excitability via GABA-A receptors.