Nan Sui

Orexins in the paraventricular nucleus of the thalamus mediate anxiety-like responses in rats

RationaleAnatomical studies have shown that the paraventricular nucleus of the thalamus (PVT) innervates areas of the forebrain involved in the expression and regulation of emotional behaviors including fear and anxiety. In addition, the PVT is densely innervated by fibers containing orexin-A (OXA) and orexin-B (OXB), peptides that are well-known for their arousal effects on behavior.ObjectivesIn this study, we investigate whether microinjections of orexin receptor agonists and antagonists in the PVT region alter expression of anxiety-like behaviors in the rat as measured in the elevated plus maze.ResultsWe report that microinjections of OXA and OXB in the PVT region elicited anxiety-like response as indicated by a reduction in open arm time and entries. In addition, OXA and OXB produced changes in ethological measures indicative of an anxiety state. Central administrations of antagonists for corticotropin releasing factor (CRF) or the opioid kappa receptors attenuated the anxiogenic effects produced by microinjections of OXA in the PVT region. We also provide evidence that endogenously released orexins act at the PVT to produce anxiety by showing that microinjections of TCSOX229, an orexin-2 receptor antagonist, in the PVT region attenuated the anxiogenic effects produced by a previous exposure to footshock stress.ConclusionsThis study indicates that endogenously released orexins act on the PVT to regulate anxiety levels through mechanisms involving the brain kappa and CRF receptors.

Stress Enables Synaptic Depression in CA1 Synapses by Acute and Chronic Morphine: Possible Mechanisms for Corticosterone on Opiate Addiction

The hippocampus, being sensitive to stress and glucocorticoids, plays significant roles in certain types of learning and memory. Therefore, the hippocampus is probably involved in the increasing drug use, drug seeking, and relapse caused by stress. We have studied the effect of stress with morphine on synaptic plasticity in the CA1 region of the hippocampus in vivo and on a delayed-escape paradigm of the Morris water maze. Our results reveal that acute stress enables long-term depression (LTD) induction by low-frequency stimulation (LFS) but acute morphine causes synaptic potentiation. Remarkably, exposure to an acute stressor reverses the effect of morphine from synaptic potentiation (∼20%) to synaptic depression (∼40%), precluding further LTD induction by LFS. The synaptic depression caused by stress with morphine is blocked either by the glucocorticoid receptor antagonist RU38486 or by the NMDA-receptor antagonist d-APV. Chronic morphine attenuates the ability of acute morphine to cause synaptic potentiation, and stress to enable LTD induction, but not the ability of stress in tandem with morphine to cause synaptic depression. Furthermore, corticosterone with morphine during the initial phase of drug use promotes later delayed-escape behavior, as indicated by the morphine-reinforced longer latencies to escape, leading to persistent morphine-seeking after withdrawal. These results suggest that hippocampal synaptic plasticity may play a significant role in the effects of stress or glucocorticoids on opiate addiction.

Effect of 5-aza-2-deoxycytidine microinjecting into hippocampus and prelimbic cortex on acquisition and retrieval of cocaine-induced place preference in C57BL/6 mice

The long lasting addiction-related abnormal memory is one of the most important foundations for relapse. DNA methylation may be a possible mechanism for persistence of such memory. Here we injected the DNA methyltransferases (DNMTs) inhibitor, 5-aza-2-deoxycytidine (5-aza) into hippocampus CA1 area and prelimbic cortex during the stages of acquisition and expression of cocaine-induced place preference in C57BL/6 mice. Results showed that in CA1 DNA methylation inhibitors could restrain acquisition but had no impact on expression of the cocaine-induced conditioned place preference (CPP). On the contrary, in prelimbic cortex, 5-aza had no effect on acquisition but blocked expression. Our results indicated that DNA methylation in hippocampus is required for learning; while DNA methylation in prelimbic cortex is necessary for memory retrieval. The present finding is consistent with the role of the hippocampus as a structure contributing to cocaine-induced memory acquisition, and prelimbic cortex, a part of prefrontal cortex as an area responsible for cocaine-induced memory retrieval. In conclusion, DNA methylation does play an important role in drug-induced learning and memory although the detailed effect still calls for further research.

Changes in emotional behavior produced by orexin microinjections in the paraventricular nucleus of the thalamus.

The paraventricular nucleus of the midline thalamus (PVT) innervates areas of the extended amygdala known to play a key role in the expression of emotional behaviors. In this study, microinjections of orexins (hypocretins), which have excitatory actions on neurons in the PVT, in the midline thalamus were used to investigate if the PVT modulates the expression of emotional behavior in the open field. First, the approach-avoidance tendency (number and duration of visit to the center area) associated with novelty was examined in orexin treated rats before and after placing a novel object in the center of the open field. Second, the expression of ethological behaviors (rearing, locomotion, freezing, and grooming) in the open field was used to determine the effects of orexins on emotionality. Microinjections of orexin-A (OXA) or orexin-B (OXB) in the PVT decreased exploration of the center area and the novel object indicating that the center area and the object had more aversive properties in orexin treated rats. Both OXA and OXB microinjections in the PVT increased the expression of freezing and grooming behaviors which are indicative of a negative emotional state. The results indicate that microinjections of orexins in the PVT made the test situation more aversive and produced avoidance behaviors. This suggests that orexins may act at the PVT to modulate behaviors associated with a negative emotional state.

Orexins in the midline thalamus are involved in the expression of conditioned place aversion to morphine withdrawal

Previous studies have implicated the bed nucleus of the stria terminalis, central nucleus of the amygdala and the shell of the nucleus accumbens (collectively called the extended amygdala) as playing an important role in mediating the aversive emotion associated with opioid withdrawal. The paraventricular nucleus of the thalamus (PVT) provides a very dense input to the extended amygdala, and the PVT is densely innervated by orexin neurons, which appear to be involved in producing some of the physical and emotional effects associated with morphine withdrawal. In the present study, we confirm that the PVT is densely innervated by orexin fibers, whereas the regions of the extended amygdala associated with the effects of morphine withdrawal are poorly innervated. Microinjections of the orexin-1 receptor (OX1R) antagonist SB334867 or the orexin-2 receptor (OX2R) antagonist TCSOX229 at doses of 5.0 or 15.0 microg into the PVT region did not affect the acquisition of the conditioned place aversion (CPA) nor the physical effects produced by naloxone-precipitated morphine withdrawal. In contrast, microinjections of TCSOX229 (15.0 microg) in the PVT region significantly attenuated the expression of naloxone-induced CPA while microinjections of SB334867 at the same dose had no effect. The results from these experiments indicate a role for OX2R in the PVT on the expression of CPA associated with morphine withdrawal. Orexins may mediate the aversive effects of morphine withdrawal by engaging the extended amygdala indirectly through the action of orexins on the PVT.

Orexin-A acts on the paraventricular nucleus of the midline thalamus to inhibit locomotor activity in rats

Orexins (hypocretins) are novel peptides that have been shown to play a role in control of behavioral arousal. The paraventricular nucleus of the midline thalamus (PVT) is one area of the brain that is the most densely innervated by orexin fibers. In addition, the PVT sends a dense projection to the nucleus accumbens, an area of the striatum involved in the regulation of locomotion. This study was done to determine the effect of microinjections of orexin-A (OXA) or the orexin receptor antagonist SB334867 in the PVT on locomotor activity (LA) in morphine-naïve and morphine-sensitized rats. Microinjections of OXA (3 microg/500 nl) in or near the PVT inhibited LA in rats tested in a novel and familiar environment as well as in rats expressing behavioral sensitization to repeated injections of morphine. In contrast, microinjections of SB334867 had no effect on LA in any of the test situations. Using an approach involving experimenter based analysis of ethological behaviors; we found that microinjections of OXA in the midline thalamus decreased LA while at the same time increasing the expression of grooming and freezing. These results suggest that OXA can act on the PVT and the midline thalamus to produce arousal independent of LA.

Morphine induces Beclin 1- and ATG5-dependent autophagy in human neuroblastoma SH-SY5Y cells and in the rat hippocampus.

Chronic exposure to morphine can induce drug addiction and neural injury, but the exact mechanism is not fully understood. Here we show that morphine induces autophagy in Neuroblastoma SH-SY5Y cells and in the rat hippocampus. Pharmacological approach shows that this effect appears to be mediated by PTX-sensitive G protein-coupled receptors signaling cascade. Morphine increases Beclin 1 expression and reduces the interaction between Beclin 1 and Bcl-2, thus releasing Beclin 1 for its pro-autophagic activity. Bcl-2 overexpression inhibits morphine-induced autophagy, whereas knockdown of Beclin 1 or knockout of ATG5 prevents morphine-induced autophagy. In addition, chronic treatment with morphine induces cell death, which is increased by autophagy inhibition through Becliln 1 RNAi. Our data are the first to reveal that Beclin 1 and ATG5 play key roles in morphine-induced autophagy, which may contribute to morphine-induced neuronal injury.

Working memory and affective decision-making in addiction: A neurocognitive comparison between heroin addicts, pathological gamblers and healthy controls

BACKGROUND Cognitive deficits are observed both in heroin dependence and in pathological gambling (PG) on various tasks. PG, as a non-substance addiction, is free of toxic consequences of drug use. Therefore a direct neurocognitive comparison of heroin addicts and pathological gamblers helps dissociate the consequences of chronic heroin use on cognitive function from the cognitive vulnerabilities that predispose addiction. METHODS A case-control design was used, comparing 58 abstinent heroin addicts, 58 pathological gamblers, and 60 healthy controls on working memory and affective decision-making functions. Working memory was assessed using the Self-ordered Pointing Test (SOPT). Affective decision-making was measured by the Iowa Gambling Task (IGT). RESULTS Heroin addicts performed significantly worse both on the IGT and on the SOPT, compared to healthy controls. Pathological gamblers performed worse on the IGT than healthy controls, but did not differ from controls on the SOPT. Years of heroin use were negatively correlated with working memory and affective decision-making performance in heroin addicts, while severity of gambling was not significantly correlated with any task performance in pathological gamblers. CONCLUSIONS Our findings indicate that deficits in affective decision-making shared by heroin dependence and PG putatively represent vulnerabilities to addiction and that working memory deficits detected only in heroin addicts may be identified as heroin-specific harmful effects.

Differential effect of NMDA receptor antagonist in the nucleus accumbens on reconsolidation of morphine -related positive and aversive memory in rats

Dysfunctional reconsolidation processes may help drug memories resist extinction and contribute to high rate of relapse. Reconsolidation of drug memory is mainly affected by the appetitive and aversive emotional experiences associated with an addictive drug. The nucleus accumbens has been shown to mediate the reconsolidation of positive emotional addictive memory, but its role in negative emotional addictive memory remains elusive. In the present study, we used morphine-induced CPP (m-CPP) and morphine-naloxone induced conditioned place aversion (m-CPA) to investigate the role of N-methyl-d-aspartate (NMDA) receptors within the nucleus accumbens on reconsolidation of emotional drug memory. Here we demonstrate that infusion of the NMDA receptor antagonist, d-(-)-2 amino-5-phosphonopentanoic acid ((D)-APV), into the nucleus accumbens before memory reactivation disrupts the reconsolidation of m-CPP, but does not affect m-CPA. The effect on m-CPP reconsolidation depended on memory reactivation: (D)-APV infusion had no effect in the absence of reactivation. The findings show that the glutamatergic NMDA receptor in nucleus accumbens mechanisms involved in reconsolidating aversive and positive morphine-associated memories can be dissociated.

Dynamic changes in orbitofrontal neuronal activity in rats during opiate administration and withdrawal

The orbitofrontal cortex is involved in the reinforcing effects of drugs of abuse. However, how the dynamic activity in OFC changes during opiate administration and withdrawal period has not been investigated. We first tested the effects of opiates and drug craving with the conditioned place preference paradigm, using manganese-enhanced magnetic resonance imaging and traditional electroencephalograph recording techniques in rats. T1-weighted 2D MRI (4.7 T) was used after unilateral injection of MnCl(2) (200 nL, 80 mM) into the right orbitofrontal cortex. The manganese-enhanced magnetic resonance imaging data suggested that the OFC activity decreased during the opiate administration period but recovered increasingly during the withdrawal period. Also, we found decreases and increases in gamma-band (20-100 Hz) activity during the opiate administration and withdrawal period, respectively. Our results showed that orbitofrontal cortex activity decreased during morphine administration and then went up progressively over several days during withdrawal. The time course of the recovery of orbitofrontal activity from inhibition during the withdrawal period may be related to the experience of drug craving.