Mecp2 truncation in male mice promotes affiliative social behavior.

Mouse models of Rett syndrome, with targeted mutations in the Mecp2 gene, show a high degree of phenotypic consistency with the clinical syndrome. In addition to severe and age-specific regression in motor and cognitive abilities, a variety of studies have demonstrated that Mecp2 mutant mice display impaired social behavior. Conversely, other studies indicate complex enhancements of social behavior in Mecp2 mutant mice. Since social behavior is a complicated accumulation of constructs, we performed a series of classic and refined social behavior tasks and revealed a relatively consistent pattern of enhanced pro-social behavior in hypomorphic Mecp2 (308/Y) mutant mice. Analyses of repetitive motor acts, and cognitive stereotypy did not reveal any profound differences due to genotype. Taken together, these results suggest that the mutations associated with Rett syndrome are not necessarily associated with autism-relevant social impairment in mice. However, this gene may be a valuable candidate for revealing basic mechanisms of affiliative behavior.

Motor and cognitive stereotypies in the BTBR T+tf/J mouse model of autism.

The BTBR T+tf/J inbred mouse strain displays a variety of persistent phenotypic alterations similar to those exhibited in autism spectrum disorders (ASDs). The unique genetic background of the BTBR strain is thought to underlie its lack of reciprocal social interactions, elevated repetitive self-directed grooming, and restricted exploratory behaviors. In order to clarify the existence, range, and mechanisms of abnormal repetitive behaviors within BTBR mice, we performed detailed analyses of the microstructure of self-grooming patterns and noted increased overall grooming, higher percentages of interruptions in grooming bouts and a concomitant decrease in the proportion of incorrect sequence transitions compared to C57BL/6J inbred mice. Analyses of active phase home-cage behavior also revealed an increase in stereotypic bar-biting behavior in the BTBR strain relative to B6 mice. Finally, in a novel object investigation task, the BTBR mice exhibited greater baseline preference for specific unfamiliar objects as well as more patterned sequences of sequential investigations of those items. These results suggest that the repetitive, stereotyped behavior patterns of BTBR mice are relatively pervasive and reflect both motor and cognitive mechanisms. Furthermore, other pre-clinical mouse models of ASDs may benefit from these more detailed analyses of stereotypic behavior.

Neural correlates of scent marking behavior in C57BL/6J mice: detection and recognition of a social stimulus.

Mice show urinary scent marking behavior as a form of social communication. Marking to a conspecific stimulus mouse or odor varies with stimulus familiarity, indicating discrimination of novel and familiar animals. This study investigated Fos immunoreactivity in inbred C57BL/6J (C57) males following scent marking behavior in response to detection of a social stimulus, or discrimination between a familiar and an unfamiliar conspecific. In Experiment 1 C57 mice were exposed for four daily trials to an empty chamber; on a test day they were exposed to the same chamber or to a male CD-1 mouse in that chamber. Increased scent marking to the CD-1 mouse was associated with increased Fos-immunoreactive cells in the basolateral amygdala, medial amygdala, and dorsal and ventral premammillary nuclei. In Experiment 2 C57 mice were habituated to a CD-1 male for 4 consecutive days and, on the 5th day, exposed to the same CD-1 male, or to a novel CD-1 male. Mice exposed to a novel CD-1 displayed a significant increase in scent marking compared to their last exposure to the familiar stimulus, indicating discrimination of the novelty of this social stimulus. Marking to the novel stimulus was associated with enhanced activation of several telencephalic, as well as hypothalamic and midbrain, structures in which activation had not been seen in the detection paradigm (Experiment 1). These included medial prefrontal and piriform cortices, and lateral septum; the paraventricular nuclei, ventromedial nuclei, and lateral area of the hypothalamus, and the ventrolateral column of the periaqueductal gray. These data suggest that a circumscribed group of structures largely concerned with olfaction is involved in detection of a conspecific olfactory stimulus, whereas discrimination of a novel vs. a familiar conspecific stimulus engages a wider range of forebrain structures encompassing higher-order processes and potentially providing an interface between cognitions and emotions.

Physical environment modulates the behavioral responses induced by chemical stimulation of dorsal periaqueductal gray in mice.

In order to investigate the relationship between behaviors elicited by chemical stimulation of the dorsal periaqueductal gray (dorsal PAG) and spontaneous defensive behaviors to a predator, the excitatory amino acid D,L-homocysteic acid (5 nmol in 0.1 micro l), was infused into the dorsal PAG and behavioral responses of mice were evaluated in two different situations, a rectangular novel chamber or the Mouse Defense Test Battery (MDTB) apparatus. During a 1-min period following drug infusion, more jumps were made in the chamber than in the MDTB runway but running time and distance traveled were significantly higher in the runway. Animals were subsequently tested using the standard MDTB procedure (anti-predator avoidance, chase and defensive threat/attack). No drug effects on these measures were significant. In a further test in the MDTB apparatus, the pathway of the mouse during peak locomotion response was blocked 3 times by the predator stimulus (anesthetized rat) to determine if the mouse would avoid contact. Ninety percent of D,L-homocysteic treated animals made direct contact with the stimulus (rat), indicating that D,L-homocysteic-induced running is not guided by relevant (here, threat) stimuli. These results indicate that running as opposed to jumping is the primary response in mice injected with D,L-homocysteic into the dorsal PAG when the environment enables flight. However, the lack of responsivity to the predator during peak locomotion suggests that D,L-homocysteic-stimulation into the dorsal PAG does not induce normal antipredator flight.

An alternative experimental procedure for studying predator-related defensive responses.

In the present study, we introduce an experimental procedure to study, in rats, a wide range of natural defensive reactions. Animals were tested in an experimental apparatus that consisted of a home cage (25 x 25 x 25 cm) connected to another chamber (25 x 25 x 25 cm-the food compartment) by a hallway (12.5 cm wide and 100 cm long, with 25-cm high walls). During 10 days before the testing procedures, each animal was isolated in the home cage, and, at the beginning of the dark phase, allowed to explore the rest of the apparatus and obtain food pellets stored in the food compartment. The testing consisted of three phases: exploring a familiar and safe environment (phase 1, on the 10th day), cat exposure (phase 2, on the 11th day), and, on the following day, exposure to the environment where the predator had been previously encountered (phase 3). These three conditions thus provided a low-defense baseline; a high level of freezing during cat exposure; and a high level of risk assessment to the hostile environment condition. An important feature of the present experimental procedure was that the behavioral responses were very stable among the animals tested within each individual phase of the testing schedule. In each phase of the testing schedule, we have also examined the Fos immunoreactivity in pontine periventricular sites related to controlling behavioral activation (i.e. the nucleus incertus) or attentional status (i.e. the locus coeruleus). Animals actively exploring a safe and familiar environment presented an increased activation of the nucleus incertus; the locus coeruleus, in turn, was particularly activated during cat exposure, and also, to lesser degree, during exposure to the hostile environment. These results give further support to the view that the animals present quite distinct behavioral states during each one of the testing situations. Taken together, the evidence suggests the present experimental procedure as particularly suitable for analyzing the neural basis of a number of specific defensive responses.

Differential effects of infralimbic vs. ventromedial orbital PFC lidocaine infusions in CD-1 mice on defensive responding in the mouse defense test battery and rat exposure test.

The ventromedial prefrontal cortex (vmPFC) is extremely sensitive to a variety of stressful situations and threatening events, and has been suggested to be an associative cortical brain system processing the integration of anxiety-related cognitive, affective and motivated behavior in rodents, primates and humans. In addition, recent evidence suggests that (a) anxiety-related affective processing appears to be lateralized to the right hemisphere vmPFC; and (b) there appears to be functional heterogeneity within the rodent vmPFC. The present study evaluated the possibility that distinct sub-areas of the right hemisphere ventral PFC might differentially influence anxiety-like defensive responding in two different predator stress situations following transient inactivation of the ventromedial orbital (vMO) or infralimbic (IL) vmPFC in CD-1 mice. In week 1, IL vmPFC lidocaine infusions reduced anxiety-like defensive responding in mice (enhanced approach and contact) confronted with a hand-held anesthetized rat stimulus in the mouse defense test battery (vMO inactivation exerted minimal effects). In week 2, vMO lidocaine infusions enhanced anxiety-like defensive responding (enhanced avoidance and protected risk assessment) toward a barricaded live rat in the rat exposure test (IL inactivation exerted minimal effects). Although it is unclear whether week 1 mouse defense test battery testing influenced week 2 rat exposure test results, these preliminary data suggest functional differences within the mouse right hemisphere ventral PFC related to cautious evaluation of predator threat. Given the dense unilateral reciprocal connectivity between the IL and vMO subregions of the PFC, both associative ventromedial cortical areas may exert complimentary yet dissociable roles in the processing of threat stimuli. This suggests that while the IL vmPFC may mediate cautious evaluation of threat situations (risk assessment), the vMO PFC may inhibit prepotent avoidance responses to facilitate such IL-mediated adaptive behavioral responses.

Infralimbic D1 receptor agonist effects on spontaneous novelty exploration and anxiety-like defensive responding in CD-1 mice.

Mesocortical dopamine (DA) terminals in the ventromedial prefrontal cortex (vmPFC) integrate cognitive/emotional processing functions underlying adaptive and appropriate behavioral responding to stressful environmental events. Results from several studies have also shown that stressor-enhanced prefrontal DA activation exerts detrimental effects on cognitive performance. However, questions have arisen as to whether stressor-enhanced vmPFC DA transmission exerts direct control over conditioned or unconditioned responses to threatening events, or whether enhanced prefrontal DA transmission gates cognitive processing to facilitate adaptive responding in threatening situations. We have previously shown that infralimbic (IL) vmPFC dopamine D2 agonist and antagonist drug infusions reduced anxiety-like responding in the elevated plus-maze (EPM) and disrupted spontaneous exploration in the Y-maze in CD-1 mice. In the present study, the effects of IL vmPFC infusions of the specific D1 receptor agonist, SKF-81297, in CD-1 mice were evaluated on spontaneous exploration in the Y-maze, anxiety-like responding in a 2-trial elevated plus-maze procedure, and anti-predator defensive responding in the Mouse Defense Test Battery (MDTB). SKF-81297 infusions disrupted spontaneous alternation performance along with potentiated repetitive 2-arm responding in the Y-maze. In the elevated plus-maze, pre-trial 1 IL SKF-81297 infusions reduced anxiety-like responding (enhanced open arm entries and time ratio, unprotected stretch attends and head dips), and reduced closed arm time ratio and protected risk assessment activity (protected stretch attends). In trial 2, 24h later (no drug infusions), open arm entries, open time ratio, and unprotected head dips remained enhanced relative to trial 2 vehicle controls. In the MDTB, avoidance distance was enhanced in the approach test; risk assessment (approach) was enhanced in the closed alley test; and defensive threat (upright postures) was enhanced in the forced contact test. Results are discussed with respect to possible influences of IL vmPFC DA receptors on cognitively mediated responding to differing levels of threat in mice.

Repeated exposure to social stress has long-term effects on indirect markers of dopaminergic activity in brain regions associated with motivated behavior.

The visible burrow system (VBS) is a chronic social stress paradigm in which a dominance hierarchy forms among male rats housed with females. Males in the VBS undergo behavioral and physiological changes thought to be manifestations of chronic social stress. Since it is unclear whether chronic social stress affects motivation and reward behavior, brain areas related to these regions were examined. Long-term effects of a single or repeated VBS exposure on mesolimbic subregions were investigated by exposing rats to the VBS either once (one cycle of VBS housing and recovery) or repeatedly (three cycles). Behavior in the VBS was observed and rats were classified as dominants or subordinates. Subordinates were further sub-classified on the basis of stress hormone (corticosterone) response to an acute stressor (i.e. restraint stress). Normal responders were categorized as stress-responsive subordinates (SRS) and animals with a blunted hypothalamic-pituitary-adrenal axis response were designated as non-responsive subordinates (NRS). Controls males were pair-housed with a single female during VBS periods and alone during recovery. Lowered enkephalin-mRNA levels were observed in the nucleus accumbens (Acb) after single VBS exposure in SRS and repeated VBS exposure both subordinate groups (i.e. SRS + NRS) compared with controls. Decreased dopamine transporter density was detected after single VBS exposure in the dorsolateral caudate putamen (DLCPu) of NRS and after repeated VBS exposure in the Acb of NRS compared with controls. Dopamine D2 receptor density was elevated after single VBS exposure in the Acb of both subordinate groups (SRS + NRS) and after repeated VBS exposure in the DLCPu, dorsomedial CPu, and Acb of NRS compared with controls. No changes in dopamine D1 receptor binding were observed in any group. These results suggest that long-term changes in dopamine activity in mesolimbic structures persist after repeated exposures to chronic social stress and may provide insight into the neurochemical basis of depressive illness and subsequent comorbidity with drug abuse vulnerability.

Development of defensive behavior and conditioning to cat odor in the rat.

Laboratory rats show a range of defensive behaviors, including freezing, avoidance, and risk assessment upon exposure to cat odor, an unconditioned but highly effective threat stimulus. This study examined defensive behaviors, and the rapid conditioning to context plus cue, of these behaviors, in 18-, 26-, and 38-day-old male and female rats exposed to cat odor. Rats were placed individually in a runway with a cloth covered (control or saturated with cat fur/skin odor) block for a 10-min trial. On the following day, a similar trial involved an odorless block. On the odor exposure day, rats of all ages showed less contact with the odor block than with the control block. The 26- and 38-day-old rats, but not the 18-day-old rats, also showed locomotor suppression, more avoidance of the area where the odor block was located, and more risk assessment than no-odor controls. On a test of conditioned behavior 24 h following exposure, 26- and 38-day-old rats exhibited defensive behavior including avoidance and reduction of locomotion while 18-day-old pups did not.

Animal models of social stress: effects on behavior and brain neurochemical systems.

Social interactions serve as an evolutionarily important source of stress, and one that is virtually ubiquitous among mammalian species. Animal models of social stress are varied, ranging from a focus on acute, intermittent, or chronic exposure involving agonistic behavior, to social isolation. The relative stressfulness of these experiences may depend on the species, sex, and age of the subjects, and subject sex also appears to influence the value of hypothalamic--pituitary--adrenal (HPA) axis activity as a general criterion for stress response: higher glucocorticoid levels are typically found in dominant females in some species. Social stress models often produce victorious and defeated, or dominant and subordinate, animals that may be compared to each other or to controls, but the appropriateness of specific types of comparisons and the interpretations of their differences may vary for the different models. Social stress strongly impacts behavior, generally reducing aggression and enhancing defensiveness, both inside and outside the stress situation. Social and sexual behaviors may be reduced in subordinate animals, as is activity and responsivity to normally rewarding events. However, some components of these changes may be dependent on the presence of a dominant, rather than representing a longer-term and general alteration in behavior. Social stress effects on brain neurotransmitter systems have been most extensively investigated, and most often found in serotonin and noradrenergic systems, with changes also reported for other monoamine and for peptidergic systems. Morphological changes and alterations of neogenesis and of cell survival particularly involving the hippocampus and dentate gyrus have been reported with severe social stress, as have longer-term changes in HPA axis functioning. These findings indicate that social stress models can provide high magnitude and appropriate stressors for research, but additionally suggest a need for caution in interpretation of the findings of these models and care in analysis of their underlying mechanisms.

Mouse defensive behaviors: pharmacological and behavioral assays for anxiety and panic.

The natural defensive behaviors of laboratory mice have been evaluated in both seminatural and highly structured situations; and characterized in terms of eliciting stimuli, response to pharmacological agents, behavior patterns, and outcome or effect on the social and physical environment. The defense patterns of laboratory mice and rats are generally similar, but mice show risk assessment on initial exposure to highly threatening stimuli while rats do not, while rats display alarm vocalizations, missing in mice. Quantitative differences in freezing and flight for laboratory mice and rats appear to largely reflect domestication effects, with wild mice and rats more similar to each other. This nexus of detailed within-species and comparative data on defense patterns makes it possible to reliably elicit specific defenses in mice or rats in an experimental context, providing well-validated assays of the natural defensive behaviors themselves, as opposed to 'models' of defense. The mouse--rat comparisons indicate considerable cross-species generality for these defense patterns, as does a scattered but considerable literature on other mammalian species, generally involving field studies and typically focusing on those aspects of defensive behavior that are visible at a distance, such as vigilance, or flight. Although potential homologies between normal mouse and human defense systems should ideally involve all four pattern components (stimulus, organismic factors, response characteristics, outcome), predictive validity in terms of response to drugs active against specific defensive psychopathology is the most extensively investigated of these. Flight, as measured in the Mouse Defense Test Battery shows a consistently appropriate response to panicolytic, panicogenic, and panic-neutral drugs, while some other predictive 'panic models' (dPAG-stimulation; DMH-inhibition; possibly conditioned suppression of drinking paradigms) also elicit and (indirectly) measure behaviors potentially related to flight. Models unrelated to flight (e.g. ultrasonic vocalization to conditioned stimuli); or for which flight elements may a relatively minor contributor to the behavior measured (Elevated T-maze) are less predictive of panicolytic or panicogenic action. These findings indicate that natural defensive behaviors provide a well-characterized pattern for analysis of effects of genetic or other physiological manipulations in the mouse, and may also serve as a model for analysis of defense-related human psychopathology.

Defense changes in stress nonresponsive subordinate males in a visible burrow system.

Dominant and subordinate male rats housed in a visible burrow system (VBS), and male controls each housed with a female, were run in a series of tests evaluating their response to threatening stimuli. Subordinates were later assessed as stress responsive (SRS) or nonresponsive (NRS) on the basis of plasma corticosterone response to restraint stress. In the tests using mildly threatening stimuli (handling, open field), NRS had longer latencies to show a righting response and reduced activity compared to SRS or (handling, open field) to dominants. There were no differences among the VBS groups in tests with more intense threat stimuli (cat odor, cat presentation). These results suggest that stressful social experience may produce a shift toward more passive and immobile forms of defense and that the experience of subordination may interact with individual differences characteristics of rats to exacerbate this shift. The hypothalamic-pituitary-adrenal (HPA) axis changes of NRS, manifested as reduced plasma corticosterone response to restraint, and previous findings of sharply reduced testosterone in these animals, along with their passive defensive behaviors, form a pattern that is suggestive of a biobehavioral "reactive" (as opposed to "proactive") coping style, and suggest that these behavioral and endocrine variables may show related changes even when altered by individual experience.

Glycogen phosphorylase reactivity in the amygdala and bed nucleus of the stria terminalis.

The present study examines the reactivity of the glial metabolic enzyme, glycogen phosphorylase, within the amygdala and bed nucleus of the stria terminalis. Reactivity for phosphorylase a, the active form of glycogen phosphorylase, was higher in all parts of the medial amygdaloid nucleus, in the medial division of the central amygdaloid nucleus, in the anterior amygdaloid area and in the bed nucleus of stria terminalis than in all parts of the lateral amygdaloid nucleus, the anterior cortical amygdaloid nucleus, the posteromedial and posterolateral cortical amygdaloid nuclei, the intercalated nucleus of the amygdala, main part and the intercalated nuclei. A greater degree of phosphorylase a reactivity was also observed in the basolateral amygdaloid nucleus, anterior and posterior parts, and in the basomedial amygdaloid nucleus, anterior part, while other parts of these nuclei were less reactive. Reactivity attributed to total glycogen phosphorylase enzyme, phosphorylase a+phosphorylase b activated by AMP, was higher and homogeneous across the amygdala. Phosphorylase a patterns are likely to reflect differences in the contribution of glycogenolysis to the metabolic support of cells in the amygdala and bed nucleus of the stria terminalis. Possible relationships to local neuronal activity and to differences in glycogenolytic neuromodulatory input are discussed.

Sexual and aggressive interactions in a visible burrow system with provisioned burrows.

The visible burrow system (VBS) is a habitat providing burrows and an open area for mixed-set rat colonies. Provisioning of food and water in the burrows makes it unnecessary for potentially defensive animals to leave the burrows to eat/drink on the surface, and enables evaluation of new types of agonistic interactions that may emerge when this necessity is removed. In such colonies, subordinate males showed high magnitude tunnel guarding behavior, occupying a tunnel opening onto the surface and confronting the dominant. Dominants, in response, made lunges into the tunnels, but quickly retreated without gaining entry, apparently stopped by contact with the defender's vibrissae. Dominants also made and continued to make lateral attacks to the wall adjacent to the tunnels guarded by subordinates, although these were useless in terms of affording contact with the subordinate. Dominant-female agonistic interactions were more frequent than those of dominants and subordinates. These were largely initiated by the male, and involved female defensive behavior. Nonetheless, females, unlike subordinates, failed to show tunnel guarding and continued to utilize the surface freely. They also spent more time in the vicinity of the dominant over days of colony formation. This apparent paradox may reflect that females were seldom wounded, and that the initial site of male contact with females was the female's anogenital area, findings suggesting that interactions of males and females often reflect male sexual advances, countered by female defenses that effectively protect nonestrus females from mounting and copulation.

Cue and context conditioning of defensive behaviors to cat odor stimuli.

Exposure of rats to a cat odor block in a previously familiarized situation was followed by three extinction days to the same or a different situation, and with or without an identical but odor-free block, and, testing in the original apparatus with an odor-free block (cue). Initial exposure produced risk assessment (stretch attend), avoidance of the block, and crouch/freeze with sniffing/head movements. Avoidance continued during extinction, but context-only exposed rats showed predominantly crouch/freeze with sniff/head movements, while rats exposed to the context+cue showed higher levels of stretch attend. During the test day, rats exposed to the cue during extinction showed reduced defensive responding compared to those not extinguished with the cue, but context extinction had less effect, possibly due in part to initial familiarization with the situation. These data indicate that both cue and context conditioning to cat odor did occur, and that the type of conditioned stimulus (context-only vs. context+cue) influenced the type of defensive behaviors elicited by this stimulus, although the all animals received the same conditioning protocol. Particular behaviors disappeared at different rates during extinction, with avoidance the most persistent. However, in this context there was no incentive for approach behaviors inconsistent with avoidance, and stretch attend behaviors could and did occur while subjects were located far from the block or the area in which it had been encountered. In addition, immobile crouch/freeze did not occur at higher than control levels, while the crouch/freeze activities that did increase incorporated sensory sampling in a relevant modality (sniffing/head movements). Thus, the behaviors seen to the conditioned stimulus appeared to reflect combinations of different defense strategies, appropriate to the type of conditioned stimulus and responsive to its extinction. Differences between these data and those from studies using fecal predator odorants suggest that the latter may not elicit a complete range of conditioned defenses.

Human defensive behaviors to threat scenarios show parallels to fear- and anxiety-related defense patterns of non-human mammals.

Defense patterns of rats and mice have been characterized in terms of the relationships between the type of defensive behavior (e.g. flight, freezing, hiding, defensive threat/attack, and risk assessment) and particular features of the eliciting (threat) stimulus and the situation in which it is encountered. Because the defense systems of rodents serve as major models for investigating and understanding both the physiology and the behavioral expression of emotional response to aversive stimuli, it is essential to evaluate whether these systems show strong parallels in human responsivity to threat. One hundred and sixty male and female undergraduate students read a set of 12 scenarios involving a present or potential threatening conspecific, and chose a primary defensive response to each. These scenarios were designed to vary features known to influence defensive responding in rodents: magnitude of threat; escapability of the situation; ambiguity of the threat stimulus; distance between the threat and the subject; presence of a hiding place. Male and female responses to the various scenarios were highly correlated, except for yell, scream, or call for help which was frequent for females, rare for males. However, a combination of this response category with 'attack' showed a highly positive (+0.96) male-female correlation, across scenarios.Correlations between manipulated (and rated) features of the threat stimulus and situation, and type of defensive behavior chosen, strongly supported a view that the patterning of defensive behavior is similar for humans and non-human mammals. Significant correlations were obtained relevant to eight specific hypotheses derived from the animal literature, with some support for two additional hypotheses (non-significant correlations averaging 0.4 or more in expected direction). While three predicted correlations were not supported in these findings, only a single significant correlation was obtained that had not been predicted on the basis of the animal literature. Although the scenario approach, and this application, have specific limitations, these results provide substantial suggestion of congruence between human and non-human mammal defense systems.

Cocaine-induced sniffing stereotypy changes in response to threat.

"Cocaine-induced stereotypies" have been extensively investigated on the basis that they may be capable of providing insights into behavioral and neurochemical mechanisms relevant to drug abuse and addiction. Recent work has indicated that cocaine enhances a number of defensive behaviors, and, that cocaine-enhanced sniffing may be a functional behavior pattern, potentially related to defense, prompting an investigation of the effects of threat stimuli on cocaine-enhanced sniffing. When behaviors of saline control rats were evaluated in their home cages (HC), or on exposure to a toy cat (TC) or real cat (RC), they showed minimal crouching in the HC; initial crouching declining over 5 days of repetitions to the TC; and continued, high-level crouching to the RC. Cocaine (30 mg/kg, IP) enhanced defensiveness in situations in which it had declined in the TC and RC groups. It also produced high-level sniffing, declining over 5 test days, in the HC; initial low-level sniffing to the TC, increasing over 5 test days; and very low levels of sniffing to the RC. These and previous data contribute to a view that cocaine enhances, but does not directly induce, defensive behaviors. They also indicate that external threat stimuli such as the RC, or initial presentation of the TC suppress sniffing, with sniffing returning as habituation to novel but not intrinsically dangerous stimuli reduces defensiveness. This view suggests that some component of "sensitization of cocaine-induced sniffing stereotypy" may reflect a release from defensiveness-mediated suppression of sniffing over repeated injection/testing as the subject becomes habituated to the injection procedure and to novel test situations.

Postnatal growth and behavioral development of mice cloned from adult cumulus cells.

Since the first successful cloning of mammals from adult somatic cells, there has been no examination of the learning or behavior of cloned offspring. The possibility of adverse effects on animals produced through adult somatic cell cloning is high because many natural biological processes are bypassed and DNA from adult cells, which presumably contain mutations, are used. In this study, we compared cloned mice produced by microinjection transfer of cumulus cell nuclei into enucleated oocytes, to control mice that were specifically generated to eliminate confounding factors that are unique to our cloning procedure. Postnatal weight gain of clones was significantly greater than that of controls. Preweaning development observations revealed that first appearance or performance of 3 out of 10 measures was delayed in cloned mice; however, results of subsequent tests of learning and memory, activity level, and motor skills were comparable for both groups. Together, these data suggest that nuclear transfer of adult somatic cell nuclei to produce cloned mice may delay the appearance of a few developmental milestones but it does not adversely affect the overall postnatal behavior of mice. In addition, this procedure may cause late onset of significantly increased body weight in cloned offspring, the cause or causes of which are being further examined.