Nucleus incertus Orexin2 receptors mediate alcohol seeking in rats.

Alcoholism is a chronic relapsing disorder and a major global health problem. Stress is a key precipitant of relapse in human alcoholics and in animal models of alcohol seeking. The brainstem nucleus incertus (NI) contains a population of relaxin-3 neurons that are highly responsive to psychological stressors; and the ascending NI relaxin-3/RXFP3 signalling system is implicated in stress-induced reinstatement of alcohol seeking. The NI receives orexinergic innervation and expresses orexin1 (OX1) and orexin2 (OX2) receptor mRNA. In alcohol-preferring (iP) rats, we examined the impact of yohimbine-induced reinstatement of alcohol seeking on orexin neuronal activation, and the effect of bilateral injections into NI of the OX1 receptor antagonist, SB-334867 (n = 16) or the OX2 receptor antagonist, TCS-OX2-29 (n = 8) on stress-induced reinstatement of alcohol seeking. We also assessed the effects of orexin-A on NI neuronal activity and the involvement of OX1 and OX2 receptors using whole cell patch-clamp recordings in rat brain slices. Yohimbine-induced reinstatement of alcohol seeking activated orexin neurons. Bilateral NI injections of TCS-OX2-29 attenuated yohimbine-induced reinstatement of alcohol seeking. In contrast, intra-NI injection of SB-334867 had no significant effect. In line with these data, orexin-A (600 nM) depolarized a majority of NI neurons recorded in coronal brain slices (18/28 cells), effects prevented by bath application of TCS-OX2-29 (10 μM), but not SB-334867 (10 μM). These data suggest an excitatory orexinergic input to NI contributes to yohimbine-induced reinstatement of alcohol seeking, predominantly via OX2 receptor signalling.

Kastman HE ，Blasiak A ，Walker L ，Siwiec M ，Krstew EV ，Gundlach AL ，Lawrence AJ ... -  《-》

Nucleus incertus corticotrophin-releasing factor 1 receptor signalling regulates alcohol seeking in rats.

Alcoholism is a chronic relapsing disorder, and stress is a key precipitant of relapse. The nucleus incertus (NI) is highly responsive to corticotrophin-releasing factor (CRF) and psychological stressors, receives a CRF innervation and expresses CRF1 and CRF2 receptor mRNA. Furthermore, the ascending NI relaxin-3 system is implicated in alcohol seeking in rats. Therefore, in alcohol-preferring rats, we examined the effect of bilateral injections into the NI of the CRF1 receptor antagonist, CP376395 or the CRF2 receptor antagonist, astressin-2B on yohimbine-induced reinstatement of alcohol seeking. Using quantitative PCR analysis of NI micropunches, we assessed the effects of chronic alcohol consumption on gene expression profiles for components of the relaxin-3 and CRF systems. Bilateral intra-NI injections of CP376395 (500 ng/0.25 µl) attenuated yohimbine-induced reinstatement of alcohol seeking. In contrast, intra-NI injections of astressin-2B (200 ng/0.25 µl) had no significant effect. In line with these data, CRF1 , but not CRF2 , receptor mRNA was upregulated in the NI following chronic ethanol intake. Relaxin family peptide 3 receptor mRNA was also increased in the NI following chronic ethanol. Our quantitative PCR analysis also identified CRF mRNA within the rat NI, and the existence of a newly identified population of CRF-containing neurons was subsequently confirmed by detection of CRF immunoreactivity in rat and mouse NI. These data suggest that NI neurons contribute to reinstatement of alcohol seeking, via an involvement of CRF1 signalling. Furthermore, chronic ethanol intake leads to neuroadaptive changes in CRF and relaxin-3 systems within rat NI.

Walker LC ，Kastman HE ，Koeleman JA ，Smith CM ，Perry CJ ，Krstew EV ，Gundlach AL ，Lawrence AJ ... -  《-》

Excitatory orexinergic innervation of rat nucleus incertus--Implications for ascending arousal, motivation and feeding control.

Blasiak A ，Siwiec M ，Grabowiecka A ，Blasiak T ，Czerw A ，Blasiak E ，Kania A ，Rajfur Z ，Lewandowski MH ，Gundlach AL ... -  《-》

Central orexin (hypocretin) 2 receptor antagonism reduces ethanol self-administration, but not cue-conditioned ethanol-seeking, in ethanol-preferring rats.

Brown RM ，Khoo SY ，Lawrence AJ 《-》

Involvement of orexin receptors within the hippocampal dentate gyrus in morphine-induced reinstatement in food-deprived rats.

The orexinergic system is found to cooperate in mediating stress-induced drug relapse. The orexinergic terminals innervate neurons of the hippocampal dentate gyrus (DG) which is a key structure in the maintenance and reinstatement of drug addiction. However, the specific contribution of intra-DG orexin receptors to stress-induced reinstatement has not been completely known. In the current study, the effects of intra-DG administration of SB334867, an orexin-1 receptor (OX1R) antagonist, and TCS OX2 29, an orexin-2 receptor (OX2R) antagonist, were investigated on the reinstatement induced by a sub-threshold dose of morphine and food deprivation (FD) stress. Adult male rats received different doses of SB334867 or TCS OX2 29 (3, 10, and 30 nM/0.5 μl DMSO 12%) bilaterally into the DG in separate groups, following the acquisition and extinction of morphine-induced conditioned place preference (CPP). Then, the reinstatement was evaluated by the 24 h FD stress and/or a sub-threshold dose of morphine (0.5 mg/kg, s.c.). CPP scores and locomotor activities were recorded during the test. The findings indicated that pre-treatment with the highest dose of SB334867 (30 nM) and two higher doses of TCS OX2 29 (10 and 30 nM) blocked the sub-threshold dose and FD stress-induced reinstatement of morphine. The effect of TCS OX2 29 on reduction of reinstatement was more pronounced than that of SB334867. It suggests a role for the orexin receptors, especially OX2R within the DG region in the stress-induced reinstatement of morphine-seeking behaviours in extinguished rats.

Pourhamzeh M ，Mozafari R ，Jamali S ，Motamedi F ，Ahadi R ，Haghparast A ... -  《-》