Involvement of Purinergic P2X4 Receptors in Alcohol Intake of High-Alcohol-Drinking (HAD) Rats

Alcohol Clin Exp Res. 2015 Oct;39(10):2022-31. doi: 10.1111/acer.12836. Epub 2015 Sep 3.

Abstract

Background: The P2X4 receptor (P2X4R) is thought to be involved in regulating alcohol-consuming behaviors, and ethanol (EtOH) has been reported to inhibit P2X4Rs. Ivermectin is an antiparasitic agent that acts as a positive allosteric modulator of the P2X4R. This study examined the effects of systemically and centrally administered ivermectin on alcohol drinking of replicate lines of high-alcohol-drinking (HAD-1/HAD-2) rats, and the effects of lentiviral-delivered short-hairpin RNAs (shRNAs) targeting P2rx4 on EtOH intake of female HAD-2 rats.

Methods: For the first experiment, adult male HAD-1 and HAD-2 rats were given 24-hour free-choice access to 15% EtOH versus water. Dose-response effects of ivermectin (1.5 to 7.5 mg/kg, intraperitoneally [i.p.]) on EtOH intake were determined; the effects of ivermectin were then examined for 2% w/v sucrose intake over 5 consecutive days. In the second experiment, female HAD-2 rats were trained to consume 15% EtOH under 2-hour limited access conditions, and dose-response effects of intracerebroventricular (ICV) administration of ivermectin (0.5 to 2.0 μg) were determined over 5 consecutive days. The third experiment determined the effects of microinfusion of a lentivirus expressing P2rx4 shRNAs into the posterior ventral tegmental area (VTA) on 24-hour EtOH free-choice drinking of female HAD-2 rats.

Results: The highest i.p. dose of ivermectin reduced alcohol drinking (30 to 45%) in both rat lines, but did not alter sucrose intake. HAD-2 rats appeared to be more sensitive than HAD-1 rats to the effects of ivermectin. ICV administration of ivermectin reduced 2-hour limited access intake (~35%) of female HAD-2 rats; knockdown of P2rx4 expression in the posterior VTA reduced 24-hour free-choice EtOH intake (~20%).

Conclusions: Overall, the results of this study support a role for P2X4Rs within the mesolimbic system in mediating alcohol-drinking behavior.

Keywords: Alcohol Drinking; High-Alcohol-Drinking Rats; Ivermectin; P2X4 Receptor; P2rx4.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alcohol Drinking / physiopathology*
  • Animals
  • Dose-Response Relationship, Drug
  • Female
  • Infusions, Intraventricular
  • Ivermectin / administration & dosage
  • Ivermectin / pharmacology
  • Male
  • Purinergic P2X Receptor Agonists / pharmacology
  • Purinergic P2X Receptor Antagonists / pharmacology
  • RNA, Small Interfering / pharmacology
  • Rats
  • Rats, Inbred Strains
  • Receptors, Purinergic P2X4 / drug effects
  • Receptors, Purinergic P2X4 / physiology*
  • Ventral Tegmental Area / drug effects

Substances

  • Purinergic P2X Receptor Agonists
  • Purinergic P2X Receptor Antagonists
  • RNA, Small Interfering
  • Receptors, Purinergic P2X4
  • Ivermectin