Differential effect of amphetamine over the corticotropin-releasing factor CRF2 receptor, the orexin OX1 receptor and the CRF2-OX1 heteroreceptor complex

Abstract

Stress is one of the factors underlying drug seeking behavior that often goes in parallel with loss of appetite. We here demonstrate that orexin 1 receptors (OX₁R) may form complexes with the corticotropin releasing factor CRF₂ receptor. Two specific features of the heteromer were a cross-antagonism and a blockade by CRF₂ of OX₁R signaling. In cells expressing one of the receptors, agonist-mediated signal transduction mechanisms were potentiated by amphetamine. Sigma 1 (σ₁) and 2 (σ₂) receptors are targets of drugs of abuse and, despite sharing a similar name, the two receptors are structurally unrelated and their physiological role is not known. We here show that σ₁ receptors interact with CRF₂ receptors and that σ₂ receptors interact with OX₁R. Moreover, we show that amphetamine effect on CRF₂ receptors was mediated by σ₁R whereas the effect on OX₁ receptors was mediated by σ₂R. Amphetamine did potentiate the negative cross-talk occurring within the CRF₂-OX₁ receptor heteromer context, likely by a macromolecular complex involving the two sigma receptors and the two GPCRs. Finally, in vivo microdialysis experiments showed that amphetamine potentiated orexin A-induced dopamine and glutamate release in the ventral tegmental area (VTA). Remarkably, the in vivo orexin A effects were blocked by a selective CRF₂R antagonist. These results show that amphetamine impacts on the OX₁R-, CRF₂R- and OX₁R/CRF₂R-mediated signaling and that cross-antagonism is instrumental for in vivo detection of GPCR heteromers. This article is part of the Special Issue entitled 'Receptor heteromers and their allosteric receptor-receptor interactions'.

Keywords: Addiction; G-protein-coupled receptor; GPCR; GPCR heteromer; Heteromers; Orexin; Sigma receptors; Stress.