The hallmark of our species is our ability to hybridize symbolic thinking with behavioral output. We began with the symmetrical hand axe around 1.7 mya and have progressed, slowly at first, then with greater rapidity, to producing increasingly more complex hybridized products. We now live in the age where our drive to hybridize has pushed us to the brink of a neuroscientific revolution, where for the first time we are in a position to willfully alter the brain and hence, our behavior and evolution. Nootropics, transcranial direct current stimulation (tDCS), transcranial magnetic stimulation (TMS), deep brain stimulation (DBS) and invasive brain mind interface (BMI) technology are allowing humans to treat previously inaccessible diseases as well as open up potential vistas for cognitive enhancement. In the future, the possibility exists for humans to hybridize with BMIs and mobile architectures. The notion of self is becoming increasingly extended. All of this to say: are we in control of our brains, or are they in control of us?

AIMS:Genetic disposition modulates the psychoactive effects of cannabis. Cannabinoids have a greater impact on brain regions that subserve anxiety in Wistar compared to Lewis strain rats. Here we aim to show that this correlates with strain differences in cannabinoid-induced anxiety-related behaviour.MAIN METHODS:Lewis and Wistar rats were administered vehicle or the synthetic cannabinoid receptor agonist, CP 55,940 (10, 25 and 50μg/kg) before testing in the conditioned ultrasonic vocalization (USV), cat odour avoidance or open area avoidance models.KEY FINDINGS:Animals were placed in a chamber in which they had previously received footshock. Wistar but not Lewis rats re-exposed under the influence of all CP 55,940 doses emitted significantly more USVs than vehicle-treated rats. In the cat odour avoidance model, rats were exposed to cat odour and given the opportunity to hide in a small box. In Wistar but not Lewis rats, 50μg/kg of CP 55,940 magnified hiding behaviour promoted by cat odour exposure. Animals were also tested in the open area avoidance model which occurred in the same arena as the predatory avoidance model but without cat odour. In Wistar, but not Lewis rats, 25 and 50μg/kg of CP 55,940 increased the avoidance of the open space.SIGNIFICANCE:CP 55,940 increased anxiety-related behaviour in Wistar rats but not Lewis rats providing a model to dissect the genetic basis of cannabinoid-induced anxiety. We show for the first time that cannabinoids magnify conditioned USVs and cat odour avoidance behaviour dependent on the strain being tested.

Individuals vary in the way in which they cope with stressful situations. It has been suggested that ‘active’ coping behaviour, characterised by aggression and territorial control, is more effective in moderating the stress associated with social defeat than ‘passive’ coping behaviour, as characterised by immobility, decreased reactivity, and low aggression. We used the rodent ‘resident/intruder’ paradigm to determine whether individual differences in coping behaviour modulate the acute adrenocortical response to social defeat. During the 10 min conflict episode, behaviours displayed by the intruder were recorded and subsequently scored. Intruders that engaged in large numbers of fights and/or frequently used physical structures to block the resident's approach (a behaviour referred to as ‘guarding’), displayed smaller corticosterone responses to defeat than other intruders. Corticosterone responses to defeat were unrelated to a measure of coping style preferences (defensive burying test) obtained prior to the defeat encounter. We further chose to investigate the neurobiological basis of this observation by comparing the patterns of defeat-induced neuronal activation in the forebrains of intruders that displayed high versus low numbers of defensive behaviours during the defeat episode. The results of this analysis indicated that ‘low fight’ and ‘low guard’ intruders, i.e. those that achieved a fight or a guard score below the 20th percentile, had significantly higher numbers of Fos-positive neurons in forebrain regions such as the medial prefrontal cortex and the amygdala than did control animals exposed to an empty resident's cage. In summary, the present data suggest that ‘active’ coping behaviour is associated with both a smaller adrenocortical response and a lower level of ‘neural activation’ following social defeat. This outcome differs from that of earlier studies, a difference that we suggest is due to the fact that the present study is the first to assess coping on the basis of behaviour actually displayed during the conflict interaction.

The automated tracking of rodents in open field environments has become a standard laboratory technique for the investigation of the effects of drugs, novel therapeutic interventions and genetic mutations on behavior. Here, we develop an extension of this technique that permits tracking in full darkness through a complex (‘enriched’) environment comprising naturalistic structures such as tunnels and hides. To eliminate unwanted light reflections and tape noise, we developed a unique video filter that combines the advantages of differential and non-differential filtering. This filter enabled the tracking of albino rats against a relatively dark background to an accuracy of approximately 97% compared to hand tracking of the same animal, irrespective of whether the rat was inside a hide box or tunnel or out in the open field. The system as a whole can be easily deployed using standard PCs and inexpensive infrared cameras and lights.

A vessel motion detection algorithm has been implemented for the measurement of multiple lymphatic chamber diameters via pre-recorded VHS tape (PAL, 25Hz). The method differs significantly from previous attempts based on edge-detection, in that it employs center of mass of pixel brightness in the vessel wall based on a Kalman filtered image subtracted from a Kalman filtered background. The operation of the algorithm confirms that it is able to detect accurately the full constriction–relaxation cycle of the vessel and produce data that meets high standards of tolerance. The procedure is well suited for real-time measurement of lymphatic and blood vessel constrictions in vitro or in vivo at multiple sites so long as the object under measurement is sufficiently differentiated from its background.

Previously we demonstrated Fos expression in the dorsal periaqueductal gray (DPAG) of the rat following cat odor exposure. Further work correlated the response to cat odor with a sustained blood pressure increase and deployment of defense behavior. It was therefore of interest to determine whether lesions of the DPAG would abolish these two effects of cat odor exposure. Male Wistar rats were given excitotoxic NMDA (N-methyl-d-aspartate) lesions of the DPAG and anterior tectum under halothane, then implanted with blood pressure telemetric probes. Sham lesions were made with saline. Rats were then exposed to cat odor with a hide option, followed 2 weeks later by re-exposure to cat odor without a hide option. Controls were exposed to rat odor in the same way. Trends toward attenuation in defense and cardiovascular indices were found in lesioned rats for cat odor exposure with a hide option, but these were not significant. Re-exposure to cat fur without a hide option enhanced the cardiovascular response and under these conditions, lesioned rats showed a significant change of the heart rate and locomotor activity response to cat fur. However, the blood pressure response was not significantly attenuated. Thus, the present results support the Fos data and indicate that the DPAG is involved in the expression of some but not all of the cardiovascular and behavioral components of the response to cat odor.

Contextual conditioned fear in the rat is characterized by a freezing immobility associated with a marked increase in blood pressure, a slow increase in heart rate, and ultrasonic vocalizations. A previous Fos study also revealed a marked activation of the ventrolateral part of the periaqueductal gray (VLPAG) and a much smaller activation of its dorsal part (DPAG). Recent chemical blockade experiments indicate that the main role of the VLPAG in the response is to impose the immobility necessary for the expression of the freezing component. We now test the role of the DPAG to see if its small activation (as revealed by Fos) is of any functional significance in the contextual fear response. Large N-methyl-D-aspartate (NMDA) excitotoxic lesions that destroyed most of the DPAG were made in 10 rats. Another group of 10 rats had sham lesions with saline. The animals were then implanted with blood pressure telemetric probes, fear conditioned, and finally tested. There was no significant difference in the amount of freezing and in the blood pressure response between the two groups. However, there was a complete abolition of ultrasonic vocalizations and a significantly greater increase in heart rate in the DPAG-lesioned group. The effect on vocalization and heart rate may be explained by lesion of adjacent structures: the lateral PAG and the superior colliculus (baroreflex alteration), respectively. Thus, most of DPAG appears to play little role in the expression of the contextual fear response.

The behavioural responses to two commonly used 'predator odours' were assessed in male Wistar rats. Cat odour was presented to rats in the form of a piece of collar that had been worn by a domestic cat. Fox odour was presented in an equivalent piece of (unworn) collar that had been impregnated with 2,4,5 Trimethylthiazoline (TMT)-an extract of fox faeces. Other rats were exposed to collars containing Triethylamine (TEA), a putrid fishy smell, or formaldehyde, which has an acrid irritating smell. Experiment 1 showed that rats approached cat odour, TMT and TEA significantly less than they did an unworn collar. However, only cat odour increased retreat to the hide box, reduced locomotor activity and elicited 'head out' behaviour. When tested immediately after odour exposure, only cat odour exposed rats showed increased anxiety in the elevated plus maze and suppressed activity in a 90-min general activity test. When returned to the odour-paired environment 24 h later in the absence of test odours, only rats that had previously received cat odour showed evidence of conditioned fear. Experiment 2 showed that rats given the benzodiazepine drug midazolam (0.5 mg/kg) display increased approach and decreased defensiveness towards a cat odour impregnated collar. In contrast, midazolam accentuated the avoidance of TMT and formaldehyde containing collars. Experiment 3 showed that when cat odour was presented in a small, enclosed environment, rats display increased body immobility, decreased grooming and increased orientation towards the odour-exuding stimulus. These responses were not seen with TMT or TEA containing collars. Taken together, these results suggest that while cat odour strongly elicits specific defensive behaviours in rats, TMT has effects that are more characteristic of an aversive odour. We suggest that the results of some previous studies using TMT may need to be reassessed.

Wistar rats were exposed to a fabric collar that had been worn by a domestic cat. Exposure took place in an open rectangular arena containing a small wooden "hide box". Rats exposed to cat odor spent more than 87% of their time in the hide box during a single 20-min exposure session, whereas rats exposed to a control odor (an unworn collar) spent less than 20% of their time hiding. One hour following this session, rats were killed and Fos immunoreactivity was compared between cat odor-exposed rats, control odor-exposed rats and an additional group that had remained in their home cages. Cat odor-exposed rats showed greater Fos expression than controls in many brain regions, particularly in the medial amygdala, medial hypothalamus and periaqueductal gray. Significant findings included strong and selective induction of Fos in the posteroventral medial amygdaloid nucleus, the premamillary nucleus (dorsal part), ventromedial hypothalamic nucleus (dorsomedial part), dorsomedial hypothalamic nucleus, periaqueductal gray (dorsomedial, dorsolateral and ventrolateral parts) and the cuneiform nucleus. Robust Fos expression in the ventromedial hypothalamus, premamillary nucleus and periaqueductal gray confirms previous suggestions of a role for these areas in predator-induced defensive behavior. Fos immunoreactivity in the medial, but not central or basolateral amygdala is a novel finding and draws attention to this subregion as a possible interface between olfactory input and emotional output.

Studies of the response of rodents to predatory odors (mainly cat) have provided useful insights into the nature of defensive behavior. This article reviews work in this area with a focus on a behavioral paradigm recently developed in our laboratory in which we present rats with a piece of fabric collar that has been previously worn by a cat. Rats presented with this stimulus spent most of their time engaged in a behavior we call 'head out' in which the rat pokes its head out from a hide box and scans the environment. Periodic 'flat back approaches' and 'vigilant rearing' towards the cat odor source are seen as well as inhibition of non-defensive behaviors such as locomotor activity and grooming. Cat odor causes a sustained increase in blood pressure (> 15mm Hg) without greatly affecting heart beat rate. Rats will develop conditioned fear to both contexts and cues that have been paired with cat odor. C-fos immunohistochemistry indicates that cat odor selectively activates a defensive behavior circuit involving the medial amygdala, ventromedial and dorsomedial hypothalamus, dorsal premammillary nucleus and the periaqueductal gray. The defensive response to cat odor is attenuated by acute administration of the benzodiazepine midazolam (0.375 mg/kg), with chronically administered SSRI antidepressants and acute alcohol exerting more modest anxiolytic effects. The behavioral response to cat odor is very different to that seen to trimethylthiazoline (TMT: fox odor) which has effects more like those seen to an aversive putrid odor. It is concluded that cat odor is a useful tool for elucidating behavioral, neural, pharmacological and autonomic aspects of defensive behavior and anxiety.

Cardiovascular and behavioral responses were recorded in rats during exposure to cat odor. Rats were habituated to an open rectangular arena that contained a small enclosed wooden box in which they could hide. On day 1 of the experiment, after 30 min in the apparatus, rats were presented with a piece of fabric collar for 60 min. On day 2, rats were presented with an identical piece of fabric collar, except that it had been worn by a cat and therefore exuded cat odor. On day 3, rats were again presented with an unworn cat collar, to determine any conditioned responses to the environment or stimulus (collar) previously associated with cat odor. Results showed significantly increased blood pressure and decreased activity during exposure to cat odor as well as avoidance of the odor stimulus and an increase in vigilance and risk-assessment measures. No significant change in heart rate was found during cat odor exposure. On day 3, a transient increase in blood pressure was seen as well as reduced activity and a range of defensive behaviors. This suggests some conditioning of fear to a context in which cat odor had previously been experienced. Heart rate was also significantly decreased on day 3. A transient rise in blood pressure was also seen when the unworn cat collar was placed into the apparatus on day 3, suggesting a conditioned response to a stimulus that has been previously associated with cat odor. This study demonstrates that a natural stressful stimulus can induce both unconditioned and conditioned autonomic and behavioral responses.

RATIONALE AND OBJECTIVES:Rodents tested in the elevated plus maze model of anxiety only show an anxiolytic response to benzodiazepines on their first exposure to the maze. The present study investigated whether a similar phenomenon occurs with benzodiazepines in a different model of anxiety that involves exposing rats to the odor of a predator.METHODS:Testing took place in a rectangular arena containing a cat odor-exuding collar at one end and a small "hide box" at the opposite end. Rats were initially familiarized with the odor-free apparatus for 20 min and then placed back in the apparatus 24 and 48 h later in the presence of cat odor.RESULTS:Vehicle-treated rats displayed marked avoidance of the cat odor on both first and second exposures, spending most of the session in the hide box and very little time near the odor source. In contrast, rats given a low dose of midazolam (0.375 mg/kg) during first exposure spent considerable time in close proximity to the odor source and much less time in the hide box. Rats given midazolam (0. 375 mg/kg) on their second exposure to cat odor displayed no such anxiolytic effect of the drug. Rats given midazolam (0.375 mg/kg) on both exposures showed a potent anxiolytic effect of the drug on each occasion. This pattern of results was replicated with a higher dose of midazolam (0.75 mg/kg). A further experiment showed that rats previously exposed to cat odor showed high levels of hiding in the test environment 24 h later even when the cat odor was no longer present. This conditioned fear was blocked by midazolam (0.75 mg/kg) suggesting that the ineffectiveness of midazolam on second exposure to cat odor is not due to a failure of the drug to affect conditioned fear.CONCLUSIONS:The ineffectiveness of midazolam in odor-experienced rats parallels the results obtained with benzodiazepines in the elevated plus maze. Such results may help illuminate the comparative lack of efficacy of benzodiazepines in treating certain types of anxiety disorders in humans.

Cat odor-induced hiding was examined in rats (Rattus norvegicus) using an apparatus with a "hide box" at one end and a piece of a worn cat collar at the other end. Rats spent most of their time hiding on exposure to the cat collar, but this response gradually habituated over repeated daily exposures. Hiding was reversed by administering the anxiolytic drug midazolam (0.375 mg/kg). Rats showed increased anxiety on the elevated plus-maze after exposure to the collar. This response was absent in habituated rats, suggesting that habituation of hiding reflects decreased odor-induced anxiety. It was established that rats located in the hide box of the apparatus might not have detected the odor of the cat collar placed at the other end of the apparatus. This implies that habituation occurs after relatively modest levels of odor exposure. Overall, the results cast some doubt on claims that predatory odors in rats are akin to phobic stimuli in humans.

Previous studies have shown that predatory odors are a potent anxiogenic stimulus for rodents, yet the ability of benzodiazepines to block odor-induced anxiety remains uncertain. The present study reevaluated this issue using a novel apparatus that, in contrast to previous studies, allowed rats to hide from the odor in a small wooden “hide box” placed within a larger arena. The odor stimulus used was a fabric cat collar that had been worn by a domestic cat for a period of 3 weeks. The experiment was divided into three phases on successive days: 1) habituation, where all rats were placed in the apparatus without cat odor present; 2) conditioning, where rats were presented with the cat odor in the apparatus; and 3) test, where rats previously exposed to the odor were tested for a conditioned avoidance response in the absence of the odor. Results showed that rats exposed to the cat collar displayed a robust avoidance response, spending about 70% of a 20-min session in the hide box compared to 25% in control rats. This avoidance response was completely reversed in rats given a low dose (0.375 mg/kg) of midazolam. During the test phase, rats exposed to the cat odor on the previous day showed elevated levels of hiding when returned to the test apparatus without the cat odor present. This conditioned avoidance was significantly attenuated in rats who had received midazolam (0.375 mg/kg) during cat odor exposure but not in rats given the same dose during the test. These results show that low-dose midazolam is an effective anxiolytic agent in rats during exposure to predatory odor.