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Reduction of circulating and selective limbic brain levels of (3α,5α)-3-hydroxy-pregnan-20-one (3α,5α-THP) following forced swim stress in C57BL/6J mice

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Abstract

Rationale

Stress activates the hypothalamic-pituitary-adrenal (HPA) axis, and GABAergic neuroactive steroids contribute to homeostatic regulation of this circuitry. Acute forced swim stress (FSS) increases plasma, cortical, and hypothalamic (3α,5α)-3-hydroxy-pregnan-20-one (3α,5α-THP) levels in rats. However, there have not been systemic investigations of acute stress on changes in plasma and brain levels of 3α,5α-THP in mouse models.

Objectives

The present experiments aimed to assess circulating and local brain levels of 3α,5α-THP following acute FSS in C57BL/6J mice.

Methods

Mice were exposed to FSS (10 min), and 50 min later, blood and brains were collected. Circulating pregnenolone and 3α,5α-THP levels were assessed in serum. Free-floating brain sections (40 μm, four to five sections/region) were immunostained and analyzed in cortical and limbic brain structures.

Results

FSS decreased circulating 3α,5α-THP (−41.6 ± 10.4 %) and reduced 3α,5α-THP immunolabeling in the paraventricular nucleus of the hypothalamus (−15.2 ± 5.7 %), lateral amygdala (LA, −31.1 ± 13.4 %), and nucleus accumbens (NAcc) shell (−31.9 ± 14.6). Within the LA, vesicular glutamate transporter 1 (VGLUT1) and vesicular GABA transporter were localized in 3α,5α-THP-positively stained cells, while in the NAcc shell, only VGLUT1 was localized in 3α,5α-THP-positively stained cells, suggesting that both glutamatergic and GABAergic cells within the LA are 3α,5α-THP-positive, while in the NAcc shell, 3α,5α-THP only localizes to glutamatergic cells.

Conclusions

The decrease in circulating and brain levels of 3α,5α-THP may be due to alterations in the biosynthesis/metabolism or changes in the regulation of the HPA axis following FSS. Changes in GABAergic neuroactive steroids in response to stress likely mediate functional adaptations in neuronal activity. This may provide a potential targeted therapeutic avenue to address maladaptive stress responsivity.

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Abbreviations

3α,5α-THP:

(3α,5α)-3-Hydroxy-pregnan-20-one

3α,5α-THDOC:

(3α,5α)-3,21-Dihydroxypregnan-20-one

3α,5β-THP:

(3α,5β)-3-Hydroxy-pregnan-20-one

5α-DHP:

5α-Dehydroxyprogesterone

3α-HSD:

3α-Hydroxysteroid dehydrogenase

BLA:

Basolateral amygdala

BNST:

Bed nucleus of the stria terminalis

CeA:

Central nucleus of the amygdala

CA1:

Cornu ammonis 1

CA3:

Cornu ammonis 3

CRF:

Corticotropin-releasing factor

DA:

Dopamine

FSS:

Forced swim stress

GC/MS:

Gas chromatography/mass spectrometry

HPA:

Hypothalamic-pituitary-adrenal

IHC:

Immunohistochemistry

LA:

Lateral amygdala

mPFC:

Medial prefrontal cortex

NAcc:

Nucleus accumbens

PVN:

Paraventricular hypothalamic nucleus

PTN:

Paraventricular thalamic nucleus

PBS:

Phosphate-buffered saline

PFA:

Paraformaldehyde

VTA:

Ventral tegmental area

VGLUT1:

Vesicular glutamate transporter 1

VGAT:

Vesicular GABA transporter

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Acknowledgments

This research was supported by the NIAAA INIA UO1-AA020935 (ALM). AMD was supported by UNC Curriculum in Toxicology NIEHS Training Grant T32 ES007126.

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Bowles Center for Alcohol Studies, University of North Carolina at Chapel Hill, 3027 Thurston Bowles Building, CB 7178, Chapel Hill, NC, 27599, USA

    Antoniette M. Maldonado-Devincci, Matthew C. Beattie, Danielle H. Morrow, Raechel E. McKinley, Jason B. Cook, Todd K. O’Buckley & A. Leslie Morrow

  2. Curriculum in Toxicology, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Antoniette M. Maldonado-Devincci & A. Leslie Morrow

  3. Curriculum in Neurobiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Jason B. Cook & A. Leslie Morrow

  4. Department of Psychiatry, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    A. Leslie Morrow

  5. Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    A. Leslie Morrow

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  1. Antoniette M. Maldonado-Devincci
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  2. Matthew C. Beattie
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Correspondence to A. Leslie Morrow.

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Maldonado-Devincci, A.M., Beattie, M.C., Morrow, D.H. et al. Reduction of circulating and selective limbic brain levels of (3α,5α)-3-hydroxy-pregnan-20-one (3α,5α-THP) following forced swim stress in C57BL/6J mice. Psychopharmacology 231, 3281–3292 (2014). https://doi.org/10.1007/s00213-014-3552-z

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