Puberty is characterized by mood swings and anxiety, which are often produced by stress. Here we show that THP (allopregnanolone), a steroid that is released as a result of stress, increases anxiety in pubertal female mice, in contrast to its anxiety-reducing effect in adults. Anxiety is regulated by GABAergic inhibition in limbic circuits. Although this inhibition is increased by THP administration before puberty and in adults, during puberty THP reduces the tonic inhibition of pyramidal cells in hippocampal region CA1, leading to increased excitability. This paradoxical effect of THP results from inhibition of α4βδ GABAA receptors. These receptors are normally expressed at very low levels, but at puberty, their expression is increased in hippocampal area CA1, where they generate outward currents. THP also decreases the outward current at recombinant α4β2δ receptors, and this effect depends on arginine 353 in the α4 subunit, a putative site for modulation by Cl−. Therefore, inhibition of α4β2δ GABAA receptors by THP provides a mechanism for the generation of anxiety at puberty.
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We thank C.A. Frye for performing steroid assays on hippocampal tissue, W. Sieghart for supplying the δ antibody, G. Homanics for supplying and genotyping the δ−/− mice, K. Perkins and D.H. Smith for discussions, and C. McBain and J. Celentano for reading the manuscript. The work in this study was supported by grants from the US National Institutes of Health (to S.S.S., C.A. and K.W.) and from the US National Institute of Alcoholism and Alcohol Abuse (to S.S.S.).
The authors declare no competing financial interests.
GABA concentration-response relationships and THP administration. (PDF 70 kb)
THP inhibition of outward current is dependent upon GABA concentration. (PDF 72 kb)
Pharmacological changes in the tonic inhibitory current of CA1 hippocampus are consistent with increased expression of α4βδ receptors after the onset of puberty. (PDF 128 kb)
THP increases the input resistance of hippocampal pyramidal cells at the onset of puberty. (PDF 30 kb)
ANOVA – F Tables – recombinant receptor experiments. (PDF 50 kb)
Mutations. (PDF 36 kb)
Tukey's test—planned post-hoc comparisons. Mutations, 1 μM GABA. (PDF 48 kb)
Tukey's test—planned post-hoc comparisons. Mutations, 10 μM GABA. (PDF 41 kb)
Western blot. (PDF 40 kb)
Slice pharmacology. (PDF 35 kb)
Tonic current. (PDF 32 kb)
Slice physiology. (PDF 32 kb)
Tukey's post-hoc comparisons. (PDF 43 kb)
Neuronal excitability. (PDF 35 kb)
Cell-attached spiking, Tukey's test—planned post-hoc comparisons. (PDF 42 kb)
Two-way ANOVA. (PDF 35 kb)
Holm-Sidak post-hoc comparison. (PDF 52 kb)
ANOVA, elevated plus maze. (PDF 33 kb)
Least significant difference planned post-hoc comparisons. (PDF 55 kb)
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