Alcohol-preferring rats show decreased corticotropin-releasing hormone-2 receptor expression and differences in hpa activation compared to alcohol-nonpreferring rats

Weidong Yong, John Paul Spence, Robert Eskay, Stephanie D. Fitz, Ruslan Damadzic, Dongbing Lai, Tatiana Foroud, Lucinda G. Carr, Anantha Shekhar, Julia A. Chester, Markus Heilig, Tiebing Liang

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background: Corticotropin-releasing hormone (CRH) and urocortins (UCNs) bind to corticotropin-releasing hormone type 2 receptor (CRF2 receptor), a Gs protein-coupled receptor that plays an important role in modulation of anxiety and stress responses. The Crhr2 gene maps to a quantitative trait locus (QTL) for alcohol preference on chromosome 4 previously identified in inbred alcohol-preferring (iP) and-nonpreferring (iNP) F2 rats. Methods: Real-time polymerase chain reaction was utilized to screen for differences in Crhr2 mRNA expression in the central nervous system (CNS) of male iP and iNP rats. DNA sequence analysis was then performed to screen for polymorphism in Crhr2 in order to identify genetic variation, and luciferase reporter assays were then applied to test their functional significance. Next, binding assays were used to determine whether this polymorphism affected CRF2 receptor binding affinity as well as CRF2 receptor density in the CNS. Finally, social interaction and corticosterone levels were measured in the P and NP rats before and after 30-minute restraint stress. Results: Crhr2 mRNA expression studies found lower levels of Crhr2 mRNA in iP rats compared to iNP rats. In addition, DNA sequencing identified polymorphisms in the promoter region, coding region, and 3'-untranslated region between the iP and iNP rats. A 7 bp insertion in the Crhr2 promoter of iP rats altered expression in vitro as measured by reporter assays, and we found that CRF2 receptor density was lower in the amygdala of iP as compared to iNP rats. Male P rats displayed decreased social interaction and significantly higher corticosterone levels directly following 30-minute restraint when compared to male NP rats. Conclusions: This study identified Crhr2 as a candidate gene of interest underlying the chromosome 4 QTL for alcohol consumption that was previously identified in the P and NP model. Crhr2 promoter polymorphism is associated with reduced mRNA expression in certain brain regions, particularly the amygdala, and lowered the density of CRF2 receptor in the amygdala of iP compared to iNP rats. Together, these differences between the animals may contribute to the drinking disparity as well as the anxiety differences of the P and NP rats.

Original languageEnglish
Pages (from-to)1275-1283
Number of pages9
JournalAlcoholism: Clinical and Experimental Research
Volume38
Issue number5
DOIs
StatePublished - 2014

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Corticotropin-Releasing Hormone Receptors
Corticotropin-Releasing Hormone
Rats
Chemical activation
Alcohols
Polymorphism
Amygdala
Assays
Messenger RNA
Chromosomes, Human, Pair 4
Quantitative Trait Loci
Neurology
Interpersonal Relations
Chromosomes
Corticosterone
DNA Sequence Analysis
Anxiety
Central Nervous System
Genes
Urocortins

Keywords

  • Alcoholism
  • Amygdala
  • Corticotropin-releasing hormone type 2 receptor
  • Selective breeding
  • Stress

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Psychiatry and Mental health
  • Toxicology
  • Medicine(all)

Cite this

Alcohol-preferring rats show decreased corticotropin-releasing hormone-2 receptor expression and differences in hpa activation compared to alcohol-nonpreferring rats. / Yong, Weidong; Spence, John Paul; Eskay, Robert; Fitz, Stephanie D.; Damadzic, Ruslan; Lai, Dongbing; Foroud, Tatiana; Carr, Lucinda G.; Shekhar, Anantha; Chester, Julia A.; Heilig, Markus; Liang, Tiebing.

In: Alcoholism: Clinical and Experimental Research, Vol. 38, No. 5, 2014, p. 1275-1283.

Research output: Contribution to journalArticle

Yong, Weidong ; Spence, John Paul ; Eskay, Robert ; Fitz, Stephanie D. ; Damadzic, Ruslan ; Lai, Dongbing ; Foroud, Tatiana ; Carr, Lucinda G. ; Shekhar, Anantha ; Chester, Julia A. ; Heilig, Markus ; Liang, Tiebing. / Alcohol-preferring rats show decreased corticotropin-releasing hormone-2 receptor expression and differences in hpa activation compared to alcohol-nonpreferring rats. In: Alcoholism: Clinical and Experimental Research. 2014 ; Vol. 38, No. 5. pp. 1275-1283.
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abstract = "Background: Corticotropin-releasing hormone (CRH) and urocortins (UCNs) bind to corticotropin-releasing hormone type 2 receptor (CRF2 receptor), a Gs protein-coupled receptor that plays an important role in modulation of anxiety and stress responses. The Crhr2 gene maps to a quantitative trait locus (QTL) for alcohol preference on chromosome 4 previously identified in inbred alcohol-preferring (iP) and-nonpreferring (iNP) F2 rats. Methods: Real-time polymerase chain reaction was utilized to screen for differences in Crhr2 mRNA expression in the central nervous system (CNS) of male iP and iNP rats. DNA sequence analysis was then performed to screen for polymorphism in Crhr2 in order to identify genetic variation, and luciferase reporter assays were then applied to test their functional significance. Next, binding assays were used to determine whether this polymorphism affected CRF2 receptor binding affinity as well as CRF2 receptor density in the CNS. Finally, social interaction and corticosterone levels were measured in the P and NP rats before and after 30-minute restraint stress. Results: Crhr2 mRNA expression studies found lower levels of Crhr2 mRNA in iP rats compared to iNP rats. In addition, DNA sequencing identified polymorphisms in the promoter region, coding region, and 3'-untranslated region between the iP and iNP rats. A 7 bp insertion in the Crhr2 promoter of iP rats altered expression in vitro as measured by reporter assays, and we found that CRF2 receptor density was lower in the amygdala of iP as compared to iNP rats. Male P rats displayed decreased social interaction and significantly higher corticosterone levels directly following 30-minute restraint when compared to male NP rats. Conclusions: This study identified Crhr2 as a candidate gene of interest underlying the chromosome 4 QTL for alcohol consumption that was previously identified in the P and NP model. Crhr2 promoter polymorphism is associated with reduced mRNA expression in certain brain regions, particularly the amygdala, and lowered the density of CRF2 receptor in the amygdala of iP compared to iNP rats. Together, these differences between the animals may contribute to the drinking disparity as well as the anxiety differences of the P and NP rats.",
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AU - Yong, Weidong

AU - Spence, John Paul

AU - Eskay, Robert

AU - Fitz, Stephanie D.

AU - Damadzic, Ruslan

AU - Lai, Dongbing

AU - Foroud, Tatiana

AU - Carr, Lucinda G.

AU - Shekhar, Anantha

AU - Chester, Julia A.

AU - Heilig, Markus

AU - Liang, Tiebing

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