Neurovascular coupling and oxygenation are decreased in hippocampus compared to neocortex because of microvascular differences.

The hippocampus is essential for spatial and episodic memory but is damaged early in Alzheimer's disease and is very sensitive to hypoxia. Understanding how it regulates its oxygen supply is therefore key for designing interventions to preserve its function. However, studies of neurovascular function in the hippocampus in vivo have been limited by its relative inaccessibility. Here we compared hippocampal and visual cortical neurovascular function in awake mice, using two photon imaging of individual neurons and vessels and measures of regional blood flow and haemoglobin oxygenation. We show that blood flow, blood oxygenation and neurovascular coupling were decreased in the hippocampus compared to neocortex, because of differences in both the vascular network and pericyte and endothelial cell function. Modelling oxygen diffusion indicates that these features of the hippocampal vasculature may restrict oxygen availability and could explain its sensitivity to damage during neurological conditions, including Alzheimer's disease, where the brain's energy supply is decreased.

Evidence that instrumental conditioning requires conscious awareness in humans.

Instrumental conditioning is a crucial substrate of adaptive behaviour, allowing individuals to selectively interact with the stimuli in their environment to maximise benefit and minimise harm. The extent to which complex forms of learning, such as instrumental conditioning, are possible without conscious awareness is a topic of considerable importance and ongoing debate. In light of recent theoretical and empirical contributions casting doubt on the early demonstrations of unconscious instrumental conditioning, we revisit the question of its feasibility in two modes of conditioning. In Experiment 1, we used trace conditioning, following a prominent paradigm (Pessiglione et al., 2008) and enhancing its sensitivity. Success in this task requires participants to learn to approach reward-predictive stimuli and avoid punishment-predictive stimuli through monetary reinforcement. All stimuli were rendered unconscious using forward-backward masking. In Experiment 2, we used delay conditioning to shorten the stimulus-outcome delay, retaining the structure of the original task but presenting the stimuli under continuous flash suppression to allow for an overlap of the stimulus, action, and outcome, as well as replacing monetary reinforcement with primary appetitive reinforcement. In both experiments, we found evidence for absence of unconscious instrumental conditioning, showing that participants were unable to learn to adjust their behaviour to approach positive stimuli and avoid negative ones. This result is consistent with evidence that unconscious stimuli fail to bring about long-term behavioural adaptations, and provides empirical evidence to support theoretical proposals that consciousness might be necessary for adaptive behaviour, where selective action is required.

Effects of serum response factor (SRF) deletion on conditioned reinforcement.

Serum response factor (SRF) is a ubiquitously expressed stimulus-dependent transcription factor that regulates gene expression by binding to serum response element in the promoter region of target genes. Recent studies in mice have shown that SRF is important for activity-dependent gene expression and synaptic plasticity in the adult brain but is dispensable for neuronal survival. Given these important functions of SRF in the CNS, it is expected to play a critical role in several aspects of learning and memory. Here we evaluated the role of SRF in conditioned reinforcement using two lines of conditional SRF mutant mice. These SRF mutant mice exhibited different spatial patterns of SRF deletion in the post-natal forebrain and notably within the hippocampus. SRF deletion was more widespread in SRF-CKCre mutants than in SRF-SynCre mutants, particularly in areas of the cortex and striatum. Mutant and wild-type mice were trained to associate one auditory cue (CS+) with reward, whereas a second cue remained relatively neutral (CS-). All mice readily acquired this discrimination, entering the food cup during CS+ but not during CS-. In a subsequent test of conditioned reinforcement, in the absence of food, wild-type control mice and SRF-SynCre mice learned to selectively perform an instrumental response that yielded CS+ presentation rather than another response that produced CS-. SRF-CKCre mutants failed to show this preferential responding for CS+. These results suggest a role for SRF in conditioned reinforcement, a manifestation of incentive learning that has been implicated in many aspects of adaptive and maladaptive behavior, such as substance abuse and eating disorders.

Neuroadaptations of total levels of adenylate cyclase, protein kinase A, tyrosine hydroxylase, cdk5 and neurofilaments in the nucleus accumbens and ventral tegmental area do not correlate with expression of sensitized or tolerant locomotor responses to cocaine.

Neuroadaptations induced by high-dose cocaine treatment have been hypothesized to persist after the cessation of drug treatment and mediate the expression of sensitization and tolerance to cocaine. We looked for evidence of these neuroadaptations in rats receiving more modest behaviorally effective cocaine treatments. Rats were exposed to either a sensitizing regimen of seven once-daily injections of 15 mg/kg cocaine or a tolerance-producing regimen involving a continuous infusion of the same daily dose. We assessed enzyme activity levels of protein kinase A and adenylate cyclase, and protein levels of tyrosine hydroxylase, cdk5 and neurofilaments in the nucleus accumbens and ventral tegmental area. Only protein kinase A activity levels were altered by cocaine treatment, but this alteration persisted for only 7 days, whereas a sensitized locomotor response was still evident at 21 days. Although behavioral tolerance to cocaine was seen the day after the termination of treatment, none of the molecular measures was altered on this or any other day. Thus, although increased protein kinase A activity can temporarily modulate sensitized responses to cocaine, alterations in total levels of the molecules assessed in our study do not correlate with the expression of sensitized or tolerant locomotor responses to cocaine.

A cannabinoid mechanism in relapse to cocaine seeking.

Treatment of cocaine addiction is hampered by high rates of relapse even after prolonged drug abstinence. This relapse to compulsive cocaine use can be triggered by re-exposure to cocaine, by re-exposure to stimuli previously associated with cocaine or by exposure to stress. In laboratory rats, similar events reinstate cocaine seeking after prolonged withdrawal periods, thus providing a model to study neuronal mechanisms underlying the relapse to cocaine. The endocannabinoid system has been implicated in a number of neuropsychiatric conditions, including drug addiction. The active ingredient of marijuana, Delta9-tetrahydrocannabinol, activates the mesolimbic dopamine (DA) reward system and has rewarding effects in preclinical models of drug abuse. We report here that the synthetic cannabinoid agonist, HU210 (ref. 13), provokes relapse to cocaine seeking after prolonged withdrawal periods. Furthermore, the selective CB1 receptor antagonist, SR141716A (ref. 14), attenuates relapse induced by re-exposure to cocaine-associated cues or cocaine itself, but not relapse induced by exposure to stress. These data reveal an important role of the cannabinoid system in the neuronal processes underlying relapse to cocaine seeking, and provide a rationale for the use of cannabinoid receptor antagonists for the prevention of relapse to cocaine use.

The ability of environmental context to facilitate psychomotor sensitization to amphetamine can be dissociated from its effect on acute drug responsiveness and on conditioned responding.

Doses of amphetamine or cocaine that fail to induce psychomotor sensitization when given to a rat in its home cage can produce robust sensitization if given immediately following placement into a relatively novel, distinct environment. A drug-associated context can serve as a conditioned stimulus, and therefore may promote robust sensitization by facilitating associative learning processes. We examined this hypothesis by habituating rats to the test environment for 1 or 6--8 hr prior to each drug injection, which degrades the ability of environmental context to serve as an effective conditioned stimulus. When 0.5 mg/kg of amphetamine was administered intravenously immediately after placement into a distinct environment there was a large acute psychomotor response (rotational behavior) on the first test day, and robust sensitization developed with repeated daily injections. When the same treatment was administered in the home cage, there was a small acute response and no sensitization developed. The enhanced acute response seen in the distinct environment was significantly attenuated by 1 hr of habituation to the test environment, and completely abolished by 6--8 hr of habituation. Also, as little as 1 hr of habituation completely prevented the development of a conditioned rotational response. In contrast, neither 1 nor 6--8 hr of habituation had any effect on the ability of amphetamine to induce robust behavioral sensitization. It is concluded that the ability of a distinct environment to facilitate sensitization to amphetamine can be dissociated from its effect on acute drug responsiveness and from the ability of drug-associated environmental stimuli to elicit a conditioned response. Possible mechanisms by which a distinct environment facilitates sensitization are discussed.

The role of contextual versus discrete drug-associated cues in promoting the induction of psychomotor sensitization to intravenous amphetamine.

The environmental context in which psychostimulant drugs are administered can have a large effect on both their acute psychomotor activating effects and their ability to induce the psychomotor sensitization associated with repeated drug administration. For example, the acute effects of amphetamine and the development of psychomotor sensitization to amphetamine and cocaine are enhanced when they are administered in a distinct and relatively novel test environment, relative to when they are given in the home cage, in the absence of any environmental stimuli predictive of drug administration. The experiments reported here were designed to further examine this phenomenon and to test the hypothesis that the ability of a distinct context to promote robust psychomotor sensitization is due to its ability to reliably signal (cue) drug administration. Specifically, we compared the ability of contextual cues (a distinct test environment) and discrete cues (light, tone and/or odor), which both reliably predict drug administration, to promote the induction of sensitization. The psychomotor stimulant effects (rotational behavior) of repeated intravenous infusions of 0. 5 mg/kg amphetamine were assessed in rats for whom drug treatments were signaled either: (1) by placement into a distinct test environment; (2) by presentation of discrete cues; or (3) rats for whom drug treatments were given in the home environment in the absence of any environmental cues. Amphetamine produced robust sensitization when given in association with placement into a distinct test environment. The same treatment failed to produce sensitization when the drug was given unsignaled in the animal's home cage. Most importantly, signaling drug administration by presenting discrete cues was not sufficient to promote the robust sensitization seen when treatments were given in a distinct test environment. These results confirm that the induction of psychomotor sensitization can be powerfully modulated by environmental context and further establish that, although contextual stimuli associated with a distinct test environment promote robust sensitization, discrete cues that merely predict drug administration do not have this property. Possible reasons for the difference in the ability of contextual versus discrete environmental cues to promote sensitization are discussed.

Susceptibility to amphetamine-induced locomotor sensitization is modulated by environmental stimuli.

We have previously reported that intravenous (i.v.) administrations of 0.5-1.0 mg/kg of amphetamine in the absence of any environmental stimuli predictive of drug administration failed to induce psychomotor sensitization whereas the same drug did produce robust sensitization when given in association with environmental novelty. These results were obtained by studying rotational behavior in animals with a unilateral 6-OHDA lesion of the mesostriatal dopamine system. The purpose of this study was to determine if environmental novelty has a similar effect on sensitization to the locomotor activating effects of amphetamine in neurologically intact rats. Rats were implanted with i.v. catheters and divided in four groups. Two groups were housed in locomotor activity cages and given seven consecutive i.v. infusions of either saline (SAL-HOME group) or 0.375 mg/kg of amphetamine (AMPH-HOME group), using a remotely activated delivery system. Simultaneously, the other two groups were transported to the test cages and given the same treatment (SAL-NOVEL and AMPH-NOVEL groups). After one week withdrawal, all groups were given an amphetamine challenge (0.375 mg/kg, i.v.). Amphetamine sensitization developed when the drug was administered under NOVEL conditions, as indicated by a progressive increase in amphetamine-induced locomotor activity over test sessions and by a greater response to the amphetamine challenge in the AMPH-pretreated versus the SAL-pretreated group. In contrast, no sensitization was observed under HOME conditions. Similar results were obtained with the analysis of vertical activity.

A comparison of two behavioral measures of psychomotor activation following intravenous amphetamine or cocaine: dose- and sensitization-dependent changes.

This paper presents data concerning the dose-effect relationships of intravenously administered amphetamine and cocaine on two widely used measures of psychomotor activation: locomotor crossover activity in neurologically intact rats, and rotational behavior in rats with a unilateral 6-hydroxydopamine lesion. There were marked differences in dose-effect relationships, both as a function of drug and of behavioral measure. Amphetamine produced a linear increase in rotational behavior over a wide range of doses (the highest effective dose was 76.8 times the lowest), but a linear increase in locomotor crossover activity over only a narrow dose range (the highest effective dose was only four times the lowest). In contrast, for cocaine, the dose-effect relationships for the two behaviors were very similar, but for both behaviors the effective dose range was quite narrow, the highest effective dose being only between two and four times the lowest. The data highlight the advantages and disadvantages of these measures as indices of the psychomotor activating effects of psychostimulant drugs.

Modulation of the induction or expression of psychostimulant sensitization by the circumstances surrounding drug administration.

The conditions necessary to induce psychomotor sensitization and to promote its expression are not well understood. Two examples are reviewed here of how the circumstances surrounding drug administration ("set and setting") can powerfully modulate the sensitization produced by psychostimulant drugs, such as amphetamine or cocaine. In the first example it is suggested that repeated exposure to psychostimulant drugs may induce "neural sensitization" (i.e., produce relevant adaptations in the nervous system). The circumstances surrounding drug administration may determine, however, whether neural sensitization is expressed in behavior. In the second example it is suggested that the circumstances surrounding drug administration may determine whether sensitization is induced at all, or at least the rate and extent of sensitization produced by a given dose of a drug. It is concluded that psychomotor sensitization is not an inevitable consequence of exposure to psychostimulant drugs, but is the result of interactions amongst the pharmacological actions of drugs and the circumstances surrounding drug administration.

Signalled versus unsignalled intravenous amphetamine: large differences in the acute psychomotor response and sensitization.

Both the acute psychomotor response and the development of sensitization to amphetamine are attenuated if i.p. injections are given in the cage where animals live (HOME), relative to when injections are given in a novel (NOVEL), but otherwise physically identical cage. It was suggested that this effect of environment on sensitization may be due to the relative availability of cues predictive of drug administration in the two conditions. It was hypothesized, therefore, that removal of all environmental cues predictive of drug administration would attenuate the development of sensitization even further. This was accomplished by comparing the psychomotor activating effects (rotational behavior) of repeated unsignalled intravenous (i.v.) infusions of 1.0 mg/kg amphetamine given in a HOME environment with those of signalled i.v. infusions given in a NOVEL environment. It was found that signalled i.v. amphetamine administration (NOVEL) produced a large acute psychomotor response, and repeated administrations resulted in a significant increase in psychomotor response (i.e., sensitization). In contrast, the same treatment in the HOME condition produced only a very small acute response and no sensitization. Indeed, the magnitude of the psychomotor response to an amphetamine challenge varied approximately 23-fold as a function of past drug history and environmental condition. It is suggested that this paradigm provides a powerful new model to study how environmental factors modulate responsiveness to psychoactive drugs.