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Injection of vehicle is not a stressor in Porsolt's swim test.

A single injection or chronic injections with vehicle are reported to induce physical changes in the brain, suggesting this treatment may be stressful. Furthermore, preliminary observations indicate that vehicle injections may interact with the behavioral effects of psychoactive compounds. We investigated the behavioral response to an injection in two tests sensitive to stress: Porsolt's swim test of depressive behavior and the resident-intruder test of aggression. Acute or repeated injections of vehicle did not alter immobility in the swim test in "normal" male NIH Swiss mice, isolated mice, or mice previously exposed to continuous fight stress. Behavior in the resident-intruder test was similar in injected and noninjected mice. The present data suggest that in male NIH Swiss mice a vehicle injection does not induced behavioral changes in tests sensitive to stress, although it is shown to alter various physiological parameters indicative of stress.

Aggression↗

Atypical anxiolytic profile of buspirone and a related drug, SM-3997, in a modified forced swim test employing straw suspension.

Previous reports have shown that immobility time increases in the presence of suspended straws in association with an inhibition of straw-climbing behavior after acute administration of a prototypical anxiolytic benzodiazepine (BZD) such as diazepam. In this modified forced swim (MFS) test employing straw suspension, the effects of two new non-BZD compounds were tested and compared with those of diazepam (0.5, 1, and 5 mg/kg, IP) used in a previous MFS test. After a 5-min test of forced swimming, four straws were suspended just above the surface of the water and subsequently the straw-climbing trials were counted for 5 min as an index of escape behaviors induced by negative emotionality (stress and/or anxiety). Rats were injected IP with either saline, buspirone HCl (0.5, 1, and 5 mg/kg), or a related compound, SM-3997 (5, 10, and 20 mg/kg), 30 min before testing. At lower doses, both buspirone (0.5, 1 mg/kg) and SM-3997 (5, 10 mg/kg) reduced the duration of immobility, as opposed to that of diazepam. Conversely, buspirone at the highest dose of 5 mg/kg significantly prolonged the duration of immobility, and SM-3997 at 20 mg/kg also prolonged its duration, indicating a biphasic effects. All doses of buspirone and SM-3997 inhibited straw-climbing counts, in the same manner as diazepam. These results suggest that buspirone may possess relatively weak and/or atypical anxiolytic effects at lower doses, whereas at 5 mg/kg this compound may have an anxiolytic effect. In addition, SM-3997 may be a less potent anxiolytic drug than buspirone in the MFS test following a single-injection protocol.

Animals↗

A new computer program for detailed off-line analysis of swimming navigation in the Morris water maze.

The program TRACK-ANALYZER runs on AT-compatible microcomputers and performs off-line analysis of data recorded from Morris water-maze experiments by means of a video tracking system or digitizing tablet. Raw data must be available on disk as ASCII-files listing position coordinates sampled at a constant frequency. Automatic recognition and correction of artifacts and missing data is a key feature of the program, as well as the option to combine commands to user-defined macros. TRACK-ANALYZER offers maximal flexibility regarding experimental schedule, maze geometry and recording parameters and may also be used to analyze open-field activity. In addition to calculating basic parameters of the swim path, such as path length and time, the program counts crossings and hits of goal platform and four virtual reference annuli, calculates search times in five different maze fields, determines directionality, tortuosity and turning preferences of swimming behavior and allows the viewing of any number of trials simultaneously on screen. ASCII-formated data output may easily be exported to commercial statistics and graphics software.

Animals↗

Mutagenic activity of swimming-pool water.

Swimming pool water, being chlorinated and exposed to trace organics from use was investigated as a possible source of mutagens using the Salmonella/mammalian-microsome test. Procedures previously described for the extraction of trace organics from water using XAD-2 macroreticular resin were modified to allow quantitative extraction of mutagens. These procedures were superior to freeze-drying and solvent-extraction. Using a base-pair histidine mutant, strain TA100, of Salmonella typhimurium significantly mutagenic responses were observed using concentrates from 3 variations of the extraction procedure. Acidified pool-water extracts eluted with ether or acetone were mutagenic, the former enhanced in the presence of the induced microsomal fraction from rat livers. Non-acidified pool-water extracts eluted with acetone were mutagenic without microsomal activation. These results indicate the presence of more than one mutagen in what is likely a complex mixture of organic molecules in swimming-pool water.

Drug Evaluation, Preclinical↗

Neuronal control of swimming locomotion: analysis of the pteropod mollusc Clione and embryos of the amphibian Xenopus.

It is rare to be able to explain the behaviour of a whole animal at the level of the properties and connections of characterized CNS neurones. In a marine mollusc, Clione, and a lower vertebrate embryo, Xenopus, it is possible to make intracellular recordings during fictive swimming behaviour. This has allowed us to analyse the operation of two central pattern generators (CPGs) at the cellular level. Although the timeframes over which the two CPGs operate are different, there are significant similarities in their patterns of neural output. A detailed analysis of the neural networks involved reveals that the swimming CPGs of Clione and Xenopus have several common operating principles, which suggests that common mechanisms have evolved to perform similar tasks, despite differences in neuronal 'hardware'.

Animals↗

Antagonistic effect of sodium ascorbate on ethanol-induced changes in swimming of mice.

Swimming behavior in the mouse was used to study the motor disturbances induced by alcohol, and the effects of vitamin C upon these disturbances were assessed. High doses of vitamin C (125 and 500 mg/kg) prevented any swimming impairment due to ethanol; however, lower doses of vitamin C (62.5 mg/kg) had no significant effect. When given 1 h before alcohol, the protective effect of vitamin C was reduced. The alcohol-induced intoxication lasted beyond alcohol's elimination from the blood, suggesting that the intoxication is maintained by a metabolite of ethanol or by an effect of ethanol, or a metabolite, on another metabolic system.

Acetaldehyde↗

Variability as a characteristic of immature motor systems: an electromyographic study of swimming in the newborn rat.

Swimming behaviour was studied in neonate rats by carrying out electromyographic recordings. The study showed that the early swimming pattern was characterized by highly instable temporal parameters. A decrease was found to occur with age in the variability of the instantaneous period in each leg and in that of the antiphase pattern. Moreover, a dissociation occurred during development between the foreleg and the backleg activity. While patterns involving the forelegs always remained extremely instable, a considerable improvement was found to occur with time in the hindlimb activity.

Afferent Pathways↗

Fast axonal transport of acetylcholinesterase in rat sciatic motoneurons is enhanced following prolonged daily running, but not following swimming.

The effects of increases in neuronal activity on fast axonal transport of acetylcholinesterase (AChE) in sciatic motoneurons were studied by subjecting rats to daily running or swimming training (8 weeks). Net accumulation of AChE activity proximal and distal to a ligature served to evaluate orthograde and retrograde transport. Results showed that runners had greater orthograde and retrograde transport of AChE as compared to control animals, while no changes were found in swimmers. These adaptations in the runners were caused by the long-term nature of the training regimen since an acute exercise session had no effect on AChE transport. The observed changes may be attributed to an increase in the mobile fraction of AChE in the motoneurons. Since swimming training had no effect on transport but entails a high level of neuronal activity, it is suggested that increased impulse activity is not the factor mediating the adaptations in axonal transport of AChE which resulted from running training.

Acetylcholinesterase↗

Apamin reduces the late afterhyperpolarization of lamprey spinal neurons, with little effect on fictive swimming.

The role of the late afterhyperpolarization (late AHP) in the firing properties of lamprey spinal neurons was tested by bath application of apamin, a selective blocker of the sk calcium-dependent potassium current. Intracellular recordings of identified motoneurons and interneurons were made with micropipette electrodes in the isolated lamprey spinal cord. Apamin reversibly reduced the amplitude of the late afterhyperpolarization without affecting other aspects of the action potential or the resting potential. The firing frequencies of the neurons were enhanced by apamin over a range of depolarizing current pulse injections. The effect of apamin was also tested on fictive swimming, which was induced in the isolated spinal cord by bath application of an excitatory amino acid (D-glutamate or N-methyl-D,L-aspartate). A concentration of apamin (10 microM) sufficient to substantially reduce the late AHP had no significant effect on the ventral root burst rate, intensity, or phase lag during fictive swimming.

Animals↗

Neonatal testosterone exposure influences neurochemistry of non-opioid swim stress-induced analgesia in adult mice.

The effects of neonatal hormone manipulations on swim stress-induced analgesia (SSIA) magnitude and neurochemical quality were examined in Swiss-Webster mice of both sexes. Previous research has indicated that non-opioid SSIA mechanisms in adult Swiss-Webster mice are sexually dimorphic. Male mice exhibit non-opioid SSIA following a 3-min swim in cold (15 degrees C) water that is antagonized by the non-competitive NMDA antagonist MK-801 (dizocilpine; 0.075 mg/kg), whereas female mice do not display NMDA-mediated analgesia in the presence of estrogen. Since male and female mice show equipotent magnitudes of SSIA, it was concluded that female mice display a neurochemically distinct, estrogen-dependent SSIA mechanism specific to their gender. In the present study, female mice exposed to testosterone during the neonatal period display NMDA-mediated analgesia even in the presence of estrogen in adulthood. Thus, expression of the female-specific, estrogen-dependent SSIA mechanism previously described may be dependent on the absence of testosterone during early ontogeny.

Analgesia↗

REM sleep deprivation decreases the antinociceptive property of enkephalinase-inhibition, morphine and cold-water-swim.

Rats treated with phosphoramidon (an enkephalinase-inhibitor 250 micrograms, i.c.v.), morphine (20 micrograms i.c.v.) or subjected to cold-water-swim (CWS, animals forced to swim in water at 5 degrees C for 5 min) showed consistent analgesia. The antinociceptive effect of phosphoramidon, morphine and CWS was antagonised by REM sleep deprivation (REMSD). It is suggested that normal duration of REM sleep is of importance for the anti-nociceptive activity of endogenous and exogenous opiates.

Analgesia↗

Impaired performance of rats in the Morris swim-maize test late in abstinence following long-term sodium barbital treatment.

Rats were tested for place learning in the Morris swim maze on days 110-114 of abstinence following 48 weeks of treatment with sodium barbital. A retarded acquisition of the swim-maze task, that could not be ascribed to motor impairments, was found in the barbital-treated rats. There was a significant difference in brain weight, but there were no significant differences between the control and barbital-treated rats in the frontal cortical concentrations of noradrenaline (NA), dopamine (DA), 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA), nor in the intra- and extrasynaptosomal activities of cerebral cortical monoamine oxidase towards NA and 5-HT. Postsynaptically, neither the cerebral cortical inositol phospholipid breakdown responses to carbachol and NA (mediated by muscarinic and alpha 1-adrenergic receptors, respectively), nor the striatal and cortical densities of muscarinic receptors labelled by [3H]quinuclidinyl benzilate [( 3H]QNB) were found significantly to be altered in the barbital-treated rats. A strong correlation between the density of striatal and cortical [3H]QNB binding sites was seen for the barbital-treated (r = 0.91) but not for the control (r = -0.05) rats. It is suggested that the deficit in performance of the barbital-treated rats in the Morris maze may be related to a cholinergic dysfunction.

Animals↗

Reduction in the level of immobilization in forced swim test and ethanol intake in rats by oxygen therapy.

Experiments replicated the previous finding that rats with high immobilization time in the forced swim test (passive rats) consumed more 15% ethanol solution in a free choice situation with tap water than rats with active behavior (active rats). Exposure of passive rats to oxygen under normal and elevated (2 ata) pressure resulted in the decrease in immobilization scores in the forced swim test as well as reduction in alcohol consumption and preference.

Alcohol Drinking↗

Prenatal cocaine and alcohol exposures affect rat behavior in a stress test (the Porsolt swim test).

Prenatal cocaine and alcohol exposures have been associated with a variety of adverse effects ranging from subtle neurobehavioral abnormalities to major malformations. In this study, we used the Porsolt swim test to assess the effects of prenatal cocaine and alcohol exposures on stress-related behavior. Pregnant Long-Evans rats were injected daily with 80 mg/kg cocaine HCl (SC) or 6.2-6.5 g/kg ethyl alcohol (PO) from gestation days 7-20 with half the dose given in the morning and the other half in the afternoon. Pair-fed and ad lib control groups were also used. One male offspring from each litter was evaluated in the Porsolt swim test at the age of 120 days. The alcohol and cocaine groups were less immobile (i.e., struggled more to escape) than the controls. These results suggest that prenatal exposure to either alcohol or cocaine can adversely affect behavior in stressful or fearful situations.

Animals↗

Size and cell number of the utricle in kinetotically swimming fish: a parabolic aircraft flight study.

Humans taking part in parabolic aircraft flights (PAFs) may suffer from space motion sickness (SMS, a kinetosis). Since it has been repeatedly shown earlier that some fish of a given batch also reveal a kinetotic behavior during PAFs (especially so-called spinning movements and looping responses) and due to the homology of the vestibular apparatus among all vertebrates, fish can be used as model systems to investigate the origin of susceptibility to motion sickness. Therefore, we examined the utricular maculae (they are responsible for the internalization of gravity in teleosteans) of fish swimming kinetotically at microgravity in comparison with animals from the same batch who swam normally. On the histological level, it was found that the total number of both sensory and supporting cells of the utricular maculae did not differ between kinetotic animals as compared to normally swimming fish. Cell density (sensory and supporting cells/100 micrometers2), however, was reduced in kinetotic animals (p<0.0001), which seemed to be due to malformed epithelial cells (increase in cell size) of the kinetotic specimens. Susceptibility to kinetoses may therefore originate in malformed sensory epithelia.

Animals↗

Deletion of the 5-HT3 receptor differentially affects behavior of males and females in the Porsolt forced swim and defensive withdrawal tests.

The central serotonin (5-HT) system is important in regulating behaviors associated with anxiety and depression. While a fair amount is known about the role of 5-HT1 and 5-HT2 receptor subtypes in regulating these behaviors, much less is known about the involvement of the 5-HT3 receptor, especially with regards to its role in sex differences in behavior. Our goal in the present studies was to examine whether deletion of the 5-HT3 receptor produces different effects in adult male and female mice on performance in three behavioral tests. We examined behavior of male and female mice lacking the 5-HT3 receptor (knock-out or KO) and their wild-type (WT) littermates in the Porsolt forced swim test because of its importance in reliably detecting anti-depressant efficacy. In addition, we examined behavior in the defensive withdrawal test and repeated exposure to an open field because behavior in these two tests provides measures of anxiety. In the Porsolt swim test, sex differences were eliminated by deletion of the 5-HT3 receptor while deletion had no effect in the habituation of locomotor activity to repeated exposure to an open field. In the defensive withdrawal test, deletion of the 5-HT3 receptor had more complex effects though these effects tended to be in the opposite direction in males and females. Together these results suggest that the 5-HT3 receptor regulates behavior-related to depression and anxiety differently in males and females. Whether these effects are due to the interaction of 5-HT3 receptor with gonadal hormones requires further examination.

Animals↗

Effects of callosal agenesis on rotational side preference of BALB/cCF mice in the free swimming test.

In order to test the hypothesis that the ontogenetic development of the corpus callosum is related to the establishment of behavioral laterality, the rotatory behavior in the free swimming test was studied in male mice of the BALB/cCF strain, in which approximately 20% of the animals present total or partial callosal agenesis. All animals were submitted to three sessions of the free-swimming rotatory test in three different sessions (diameter of the recipient = 21 cm; session duration = 5 min; inter-test interval = 48 h). The number and direction of the 30 and 360 degrees turns were recorded. Animals were classified as side-consistent turners (to the right or to the left) when they did not change their preferred side of rotation in all three sessions and in both turning units. In general our results suggested that acallosal animals present more pronounced laterality than normal ones. In the acallosal group, the percentage of consistent turners was significantly higher than that of non-consistent turners. The percentage of animals that presented strong turning preferences in the acallosal group was higher than in the normal group. In first session, the acallosal group presented a higher average number of turns to preferred side than the normal group. Taken together, our results constitute an endorsement to the hypothesis that the normal development of the corpus callosum is related to the establishment of cerebral laterality.

Agenesis of Corpus Callosum↗

Behavioral responses during the forced swim test are not affected by anti-inflammatory agents or acute illness induced by lipopolysaccharide.

Pro-inflammatory cytokines and other molecules traditionally associated with immune function have been implicated in mediating behavioral and physiological consequences of stressor exposure. There is also evidence that cytokines are aberrantly expressed in depressive populations, suggesting they may play an etiological role in the development of depression/despair-related processes. Thus, we conducted a series of experiments to determine whether agents known to suppress cytokine activity or inflammatory responses in the CNS would alter the normal progression of behavioral responses during the forced swim test (FST, an animal model of depression/behavioral despair). Adult male Sprague-Dawley rats were injected with indomethacin (1 or 10 mg/kg intraperitoneally (i.p.)), alpha-MSH (0.25 or 0.5 microg icv), or minocycline (20 or 40 mg/kg i.p.) prior to each day of the FST and behavioral assessments were performed. Injection of indomethacin, alpha-MSH, or minocycline had no effect on the development of the immobility response during the FST on either day of testing. In a second series of experiments, we examined whether behavioral responses during forced swim would be affected by acute illness induced by a single injection of lipopolysaccharide (LPS). Acute injection of LPS (10 or 100 microg/kg i.p.) had no effect on behavioral responding during the FST irrespective of when it was injected, despite pronounced reductions in social behavior following these same doses of LPS. From these studies, we conclude that (a) endogenous inflammatory mediators do not appear to be involved in the normal progression of behavioral responses during the FST, and (b) behavioral responses during the FST are not affected by acute systemic injection of LPS.

Analysis of Variance↗

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