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Effects of a stressor and corticotrophin releasing factor on ethanol deprivation-induced ethanol intake and anxiety-like behavior in alcohol-preferring P rats

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Abstract

Rationale

Stress may elevate ethanol drinking and anxiety associated with ethanol drinking. Studies to identify relevant neurobiological substrates are needed.

Objective

To assess roles of brain regions in corticotrophin releasing factor (CRF) effects on stressor-enhanced, ethanol deprivation-induced drinking and anxiety-like behavior.

Methods

Ethanol-preferring rats (P rats) were exposed to three cycles of a two-bottle choice paradigm with two 2-day deprivation periods that included 1 h exposure to a restraint stressor. To assess the role of CRF and to identify relevant brain regions, a CRF-1 receptor antagonist (SSR125543; 10 ug) was injected into the nucleus accumbens (NAC), amygdala (Amyg), or dorsal raphe nucleus (DRN) prior to exposure to the restraint stressor. In a second study, CRF (0.5 ug) was injected into one of these regions, or the ventral tegmental area (VTA), or paraventricular nucleus of the hypothalamus (PVN).

Results

Applying the restraint stressor during deprivation increased voluntary intake and sensitized anxiety-like behavior. Antagonist injection into the NAC prevented increased drinking without affecting anxiety-like behavior, whereas injection into the Amyg or DRN prevented the anxiety-like behavior without affecting drinking. To confirm CRF actions in the stressor effect, CRF was injected into selected brain regions. NAC injections (but not the VTA, Amyg, DRN, or PVN) facilitated drinking but did not change anxiety-like behavior. Injections into the DRN or Amyg (but not PVN or VTA) enhanced anxiety-like behavior.

Conclusions

Results emphasize that a restraint stressor elevates ethanol intake and sensitizes ethanol deprivation-induced anxiety-like behavior through CRF1 receptors in the NAC and Amyg/DRN, respectively.

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Acknowledgments

This work was supported by NIAAA [AA11605, AA14949] as well as by funds from the Center for Alcohol Studies. Drs. Knapp and Overstreet contributed equally to this work. We thank T-K Li and Lawrence Lumeng at Indiana University Medical Center for sending the original breeding stock to establish our colony of P rats used in this investigation. We also thank Kui-Ling Huang for technical assistance. The authors confirm that they have no financial relationship with the NIH or Synofi other than funding this research (NIH) or providing the gift of the SSR compound (Synofi). The authors also confirm that these experiments comply with all relevant laws of the United States.

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Authors and Affiliations

  1. Bowles Center for Alcohol Studies, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27514-7178, USA

    Darin J. Knapp, David H. Overstreet, Mae Huang, Tiffany A. Wills, Buddy A. Whitman, Robert A. Angel, Sarah E. Sinnett & George R. Breese

  2. Neurobiology Curriculum, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27514-7178, USA

    Darin J. Knapp, Tiffany A. Wills, Buddy A. Whitman & George R. Breese

  3. Neuroscience Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27514-7178, USA

    George R. Breese

  4. Department of Psychiatry, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27514-7178, USA

    Darin J. Knapp, David H. Overstreet & George R. Breese

  5. Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27514-7178, USA

    Sarah E. Sinnett & George R. Breese

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  1. Darin J. Knapp
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  2. David H. Overstreet
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Correspondence to Darin J. Knapp.

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213_2011_2366_MOESM1_ESM.ppt

Supplemental Figure Legend. Microinjection targeting based on the brain atlas of Paxinos and Watson (2005). See “Methods” in text for coordinate details for each brain region. Red circles represent the ideal center point (anterior-posterior, dorso-ventral, and medial-lateral) based on the coordinates detailed in Table 1. For the schematics at left, dark hatched circles represent approximate outer limits of injections deemed acceptable. Other circles indicate representative locations of injections in regions of experimental groups in which CRF (green circles) or the CRF antagonist (black circles) were active. Numbers in the lower right of each schematic reflect the plate number from the atlas. Photomicrographs representing injection sites for each region appear in the right column. The straight hatched lines in the photomicrographs represent the approximate angle of approach to the sites of interest where darkened dye markings can be seen. a Accumbens, b paraventricular nucleus of the hypothalamus, c amygdala, d ventral tegmental area, e dorsal raphe nucleus (PPT 2.04 mb)

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Knapp, D.J., Overstreet, D.H., Huang, M. et al. Effects of a stressor and corticotrophin releasing factor on ethanol deprivation-induced ethanol intake and anxiety-like behavior in alcohol-preferring P rats. Psychopharmacology 218, 179–189 (2011). https://doi.org/10.1007/s00213-011-2366-5

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