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Mouse Testing Methods in Psychoneuroimmunology: Measuring Behavioral Responses.

The field of psychoneuroimmunology (PNI) aims to uncover the processes and consequences of nervous, immune, and endocrine system relationships. Behavior is a consequence of such interactions and manifests from a complex interweave of factors including immune-to-neural and neural-to-immune communication. Often the signaling molecules involved during a particular episode of neuroimmune activation are not known, but behavioral response provides evidence that bioactives such as neurotransmitters and cytokines are perturbed. Immunobehavioral phenotyping is a first-line approach when examining the neuroimmune system and its reaction to immune stimulation or suppression. Behavioral response is significantly more sensitive than direct measurement of a single specific bioactive and can quickly and efficiently rule in or out relevance of a particular immune challenge or therapeutic to neuroimmunity. Classically, immunobehavioral research was focused on sickness symptoms related to bacterial infection, but neuroimmune activation is now a recognized complication of diseases and disorders ranging from cancer to diabesity to Alzheimer's. Immunobehaviors include lethargy, loss of appetite, and disinterest in social activity/surrounding environment. In addition, neuroimmune activation can diminish physical activity, precipitate feelings of depression and anxiety, and impair cognitive and executive function. Provided is a detailed overview of behavioral tests frequently used to examine neuroimmune activation in mice with a special emphasis on pre-experimental conditions that can confound or prevent successful immunobehavioral experimentation.

Animals

Physiological and behavioral responses to starvation in the golden hamster.

Physiological and behavioral responses of adult hamsters to starvation were studied by measuring food intake, weight recovery, serum concentrations of glucose, insulin, free fatty acids and beta-hydroxybutyrate, and ketonuria in animals subjected to different weight losses, diets, and durations of fast. Hamsters were debilitated by fasts longer than 12 h or leading to greater than 20% weight loss. Hamsters' feeding patterns were unmodified by fasts ranging between 5 and 12 h and showed no circadian periodicity. Hamsters predominantly recovered from weight losses without increasing their food consumption (unless they were offered a diet of pellets and seeds) and without changing their meal patterns, at a rate of weight gain proportional to the magnitude of preceding weight loss if provided with uninterrupted access to food. By 8 h of fast, blood metabolites were indicative of mobilization of body fat. Hamsters are thus behaviorally unresponsive to duration of fast, but compensate physiologically for weight losses with proportional increases in the rate of weight gain.

Animals

Effects of psychotropic drugs on pressor and behavioral responses to brain stimulation in unrestrained, unanesthetized rats.

Electrical stimulation of the posterior hypothalamus (PH) and the mesencephalic reticular formation (MRF) in unanesthetized, unrestrained rats with chronic electrode implants and an arterial cannula elicited a rise in blood pressure accompanied by behavioral changes such as exploration, flight or escape responses. Pentobarbital inhibited both the pressor and behavioral responses to PH and MRF stimulation. Chlorpormazine, diazepam and imipramine depressed the pressor response to PH stimulation rather than that to MRF stimulation with affecting the behavioral responses. It is concluded from these results that chlorpromazine, diazepam and imipramine exert their action on the neural pathway involved in the pressor response rather than on that inducing behavioral responses, whereas pentobarbital affects more extended brain areas related to these neural systems.

Animals

Behavioral response to vasectomy.

This article reviews the literature concerning the behavioral and psychological response of men and their wives to the vasectomy operation when used as a contraceptive technique. The data suggest that two primary changes may frequently be observed postoperatively. Individuals who have experienced anxiety in conjunction with intercourse because of a fear of contraceptive failure or pregnancy tend to report a decline in anxiety and a disinhibition of sexual arousal postoperatively. Also, test results suggest that behavoir patterns within the marital dyad are altered in some cases when the men adopt stereotyped masculine behavior, presumably to deny any suggestion that they are less masculine because of the operation. Ramifications of these findings as well as numerous other speculations, are considered, along with the suggestion that prescreening and postvasectomy counseling may help reduce negative response to the operation.

Adult

Development and degeneration of hearing in the C57/b16 mouse: relation of electrophysiologic responses from the round window and cochlear nucleus to cochlear anatomy and behavioral responses.

In disorders of the auditory organ associated with hearing loss, a correlation of the anatomical, behavioral and electrophysiological studies furnishes the investigator several parameters of study to better comprehend the hearing mechanism and the various anatomical structural dysfunctions that relate to the hearing loss. Mice are interesting to study because they carry several types of genes for hereditary deafness. Until recently, it was very difficult to train mice for behavioral hearing threshold testing; however, having developed such a method, the next step was to study the normal mice in these three parameters and extend the studies to mice with hereditary hearing loss. The type of mouse studied in this research was the C57/b16 strain which carries such a gene. The results of the studies indicate that the results of the behavioral and electrophysiological studies relate well to each other, and that light microscopic studies (if taken alone) do not furnish us with the actual functional capability of the structures of the inner ear. It is recommended (and it would be most valuable) that efforts should be made to collect and study human temporal bones with all three parameters of study for a better understanding of the structures of the human inner ear. Reference is made and recommendations given as to the parameters of studies while performing human electrocochleography.

Acoustic Stimulation

Different behavioral responses to L-DOPA after anterolateral or posterolateral hypothalamic injections of 6-hydroxydopamine.

After bilateral microinjections of 6-hydroxydopamine (6-OHDA) into the anterolateral (AL) or posterolateral (PL) hypothalamus L-DOPA (1,3,4-dihydroxy-phenylalanine) produced running and rearing in AL6-OHDA rats and oral stereotypies in PL 6-OHDA rats. Since the same dose of L-DOPA had no behavioral effect in vehicle injected rats, the responses to L-DOPA of both AL and PL 6-OHDA rats are examples of behavioral supersensitivity. The different forms of behavioral supersensitivity correlated with different patterns of catecholamine (CA) denervation determined by fluorescent microscopy. The major regions of CA denervation in AL 6-OHDA rats were neocortex, hippocampus, limbic forebrain, anteromedial striatum and anterolateral hypothalamus. PL 6-OHDA had these same areas denervated and, in addition, had severe denervation of the entire striatum, parts of the amygdala and thalamus, and of the posterolateral hypothalamus. We conclude that the supersensitive behavioral response to a fixed dose of L-DOPA is determined by the pattern and/or extent of CA denervation.

Afferent Pathways

[Relationship between the incremental and behavioral responses evoked by stimulation of the ventral anterior nucleus of the rat thalamus].

Low frequency stimulation of nucleus Ventralis Anterior thalami in unanesthetized rats elicits incremental responses and produces, in some conditions, characteristic behavioral reaction. Stimulation thresholds giving rise to behavioral reaction are higher than those allowing to find electrocortical responses. The analysis of their evolution discloses changes which are bound to both of the following parameters: the stimulation frequency and the arousal. The lowest behavioral thresholds are got at 8-10 c/s during relaxed wakefulness whereas the lowest electrocortical thresholds are found at 5-8 c/s during slow-wave sleep.

Animals

[Inhibition of the behavioral responses of cats after discontinuing electric stimulation of the caudate nucleus].

A prolonged suppression of behavior occurs in free-moving cats after disruption of low-frequency stimulation of the caudate nucleus head. Its outward manifestations and EEG-pattern look like those of natural sleep. However, unlike sleep, during caudate suppression motor functions are more affected, the activation response to presented test-stimuli is shorter, and the emotionality of animals is weaker. The suppression is most pronounced following the stimulation of rostro-medio-dorsal parts of the nucleus head. In small doses the neuroleptic haloperidol enhances, and the psychostimulant d,l-amphetamine decrease the caudate effect. It is suggested that the trace response triggered by the stimulation of the caudate nucleus is closer to behavioral suppression than to sleep and that it depends on the activity of the dopaminergic mechanisms.

Amphetamine

Inversion of a behavioral response in bacterial chemotaxis: explanation at the molecular level.

Certain cheU mutants of Salmonella show inverted chemotactic behavior, being repelled by attractants and attracted by repellents. Such a dramatic change in behavioral pattern would seem at first glance to require drastic and complex alterations in the sensory processing system. In fact, the behavior can be explained by a simple shift in the level of a response regulator and the subtle effects of this shift on flagellar function. Flagella can exist in either a left-handed or a right-handed structure depending on applied torsion. Wild-type cells swim smoothly by counterclockwise rotation of a left-handed helical bundle and tumble when the motors briefly reverse to clockwise rotation (normal random motility). The cheU mutation causes a shift in response regulator level relative to the critical threshold value, resulting in extended clockwise operation so that the flagella are fully converted to the right-handed helical form. These cells therefore swim smoothly by clockwise rotation of a right-handed bundle and tumble when the motor briefly reverses to counterclockwise rotation (inverse random motility). Thus, tumbling is associated with brief reversals and not with a particular sense of rotation. A wild-type cell, with its steady-state response regulator level placing it initially in normal random motility, will swim smoothly on addition of attractant, whereas a cheU mutant with inverse random motility will tumble given the same stimulus. The phenomenon illustrates the profound behavioral consequences that can result from a single mutation in a key gene.

Chemotaxis

Social status in zebrafish modulates the behavioral response to 5-HT2C receptor agonists and antagonists.

The effects of previous social experiences on social behavior have been demonstrated across species both in cooperative and competitive contexts. In dominance-subordinate hierarchies, differences across social ranks have been observed in many different mechanisms. Dominance hierarchies interfere in defensive behavior, where subordinate animals present a greater defensive behavior, regarding potential threats ("anxiety-like behavior"), than dominant animals. The serotonergic system plays a key role in regulating and mediating threat responses, including 5-HT2 receptors in the types of proximal threat responses modulated by the stress of social defeat. We separated 148 adult zebrafish in pairs and allowed them to interact for five days; after that, the dominant-subordinate rank was determined, and animals were treated with a 5-HT2C receptor agonist (MK-212) or antagonist (RS-102221) before being observed in the novel tank test. While MK-212 increased bottom-dwelling, erratic swimming, and freezing across all statuses, RS-102221 decreased these variables in dominants but increased them in subordinates. Moreover, the effects of MK-212 were larger in subordinates than in controls or dominants, suggesting a sensitization of the 5-HT2C receptor.

Animals

Genetic analysis of the behavioral response to d-amphetamine in mice.

The ambulatory and rearing responses to d-amphetamine were studied in a battery of recombinant inbred strains and in three closely related strains: C57BL/6J, C57BL/10J, and C57BL/6By. Differences in the increase of ambulation (stimulation) caused by d-amphetamine were seen between C57BL/6By and the other two C57BL strains. Analysis of F1 and backcross matings suggests a one-gene model. A mutation at the genetic locus that affects the response to d-amphetamine seems to have taken place in the C57BL/6By strain. Strain differences in the decrease of rearing behavior (inhibition) produced by the drug were observed in recombinant inbred strains. Although the genetic analysis is not conclusive, it appears to be compatible with regulation by a single major gene. The two single-gene models reported here (one affecting the stimulatory response and the other the inhibitory response to d-amphetamine) may be useful in the study of neural mechanisms involved in stimulation and inhibition of behavior by d-amphetamine.

5-Hydroxytryptophan

Genetic analysis of the behavioral response to d-amphetamine in mice.

The ambulatory and rearing responses to d-amphetamine were studied in a battery of recombinant inbred strains and in three closely related strains: C57BL/6J, C57Bl/10J, and C57BL/LBy. Differences in the increase of ambulation (stimulation) caused by d-amphetamine were seen between C57BL/6By and the other two C57BL strains. Analysis of F1 and backcross matings suggests a one-gene model. A mutation at the genetic locus that affects the response to d-amphetamine seems to have taken place in the C5BL/6By strain. Strain differences in the decrease of rearing behavior (inhibition) produced by the drug were observed in recombinant inbred strains. Although th of l-dihydroxyphenylalanine (L-dopa) and d,l,5-hydroxytryptophan (d,l,5-HTP) on reserpine-induced amnesia.

Animals