A pair of ascending neurons in the subesophageal zone mediates aversive sensory inputs-evoked backward locomotion in Drosophila larvae.

Animals typically avoid unwanted situations with stereotyped escape behavior. For instance, Drosophila larvae often escape from aversive stimuli to the head, such as mechanical stimuli and blue light irradiation, by backward locomotion. Responses to these aversive stimuli are mediated by a variety of sensory neurons including mechanosensory class III da (C3da) sensory neurons and blue-light responsive class IV da (C4da) sensory neurons and Bolwig's organ (BO). How these distinct sensory pathways evoke backward locomotion at the circuit level is still incompletely understood. Here we show that a pair of cholinergic neurons in the subesophageal zone, designated AMBs, evoke robust backward locomotion upon optogenetic activation. Anatomical and functional analysis shows that AMBs act upstream of MDNs, the command-like neurons for backward locomotion. Further functional analysis indicates that AMBs preferentially convey aversive blue light information from C4da neurons to MDNs to elicit backward locomotion, whereas aversive information from BO converges on MDNs through AMB-independent pathways. We also found that, unlike in adult flies, MDNs are dispensable for the dead end-evoked backward locomotion in larvae. Our findings thus reveal the neural circuits by which two distinct blue light-sensing pathways converge on the command-like neurons to evoke robust backward locomotion, and suggest that distinct but partially redundant neural circuits including the command-like neurons might be utilized to drive backward locomotion in response to different sensory stimuli as well as in adults and larvae.

Circuits That Mediate Expression of Signaled Active Avoidance Converge in the Pedunculopontine Tegmentum.

An innocuous sensory stimulus that reliably signals an upcoming aversive event can be conditioned to elicit locomotion to a safe location before the aversive outcome ensues. The neural circuits that mediate the expression of this signaled locomotor action, known as signaled active avoidance, have not been identified. While exploring sensorimotor midbrain circuits in mice of either sex, we found that excitation of GABAergic cells in the substantia nigra pars reticulata blocks signaled active avoidance by inhibiting cells in the pedunculopontine tegmental nucleus (PPT), not by inhibiting cells in the superior colliculus or thalamus. Direct inhibition of putative-glutamatergic PPT cells, excitation of GABAergic PPT cells, or excitation of GABAergic afferents in PPT, abolish signaled active avoidance. Conversely, excitation of putative-glutamatergic PPT cells, or inhibition of GABAergic PPT cells, can be tuned to drive avoidance responses. The PPT is an essential junction for the expression of signaled active avoidance gated by nigral and other synaptic afferents.SIGNIFICANCE STATEMENT When a harmful situation is signaled by a sensory stimulus (e.g., street light), subjects typically learn to respond with active or passive avoidance responses that circumvent the threat. During signaled active avoidance behavior, subjects move away to avoid a threat signaled by a preceding innocuous stimulus. We identified a part of the midbrain essential to process the signal and avoid the threat. Inhibition of neurons in this area eliminates avoidance responses to the signal but preserves escape responses caused by presentation of the threat. The results highlight an essential part of the neural circuits that mediate signaled active avoidance behavior.

Long-term UVB exposure promotes predator-inspection behaviour in a fish.

Ultraviolet-B radiation (UVB) reaching the earth's surface has increased due to human-caused stratospheric ozone depletion. Whereas the harmful effects of UVB on aquatic organisms are well studied at the molecular and cellular level, recent studies have also begun to address behavioural changes caused by sublethal amounts of UVB. However, the behavioural consequences of long-term exposure to ecologically relevant UVB levels over several life stages are virtually unknown, particularly with regard to predator-prey behaviour. We found increased predator-inspection behaviour together with a smaller body length in three-spined sticklebacks (Gasterosteus aculeatus) after fish were exposed for about seven months to natural sunlight conditions with enhanced UVB, compared with full siblings exposed to natural sunlight only. The observed change in antipredator behaviour may reflect a direct behavioural response mediated through UVB-induced oxidative stress during development. Alternatively, the smaller body size in UVB-exposed fish may result in an increased inspection effort allowing them to spend more time foraging. Our findings suggest that, within the scope of environmental change, UVB radiation constitutes an important stress factor by eliciting behavioural responses that influence crucial ecological processes, such as predator-prey interactions.

Crickets alter wind-elicited escape strategies depending on acoustic context.

Acoustic signals trigger various behaviours in insects such as courtship or escape from predators. However, it remains unknown whether insects utilize acoustic signals to recognize environmental contexts. The cricket is a prominent model insect for neuroethological studies on acoustic behaviour because female crickets exhibit positive phonotaxis in response to male calling songs, and flying crickets display avoidance behaviour for high-frequency sounds such as echolocation call of bats. The carrier frequency of these sounds is a major factor in determining whether they initiate these acoustic behaviours. Here, we examined the impacts of different frequencies of tone sounds on cercal-mediated escape behaviour, using a 5-kHz tone corresponding to the calling song and a 15-kHz tone serving as a trigger of avoidance behaviours. Neither frequency elicited a response in the standing cricket by itself, but they had different impacts on walking responses to airflow stimuli. While the 15-kHz tone reduced response probability, extended moving distance, and enhanced turn-angle variability, the 5-kHz tone had no effect. Although both frequencies of tones facilitated walking backward, the 15-kHz tone had a larger effect than the 5-kHz tone. These frequency dependencies of behavioural modulation suggest that crickets can recognize acoustic contexts and alter their escape strategy accordingly.

CRF-like receptor SEB-3 in sex-common interneurons potentiates stress handling and reproductive drive in C. elegans.

Environmental conditions can modulate innate behaviours. Although male Caenorhabditis elegans copulation can be perturbed in the presence of stress, the mechanisms underlying its decision to sustain copulation are unclear. Here we describe a mating interference assay, which quantifies the persistence of male C. elegans copulation in noxious blue light. We show that between copulations, the male escapes from blue light illumination at intensities over 370 µW mm(-2). This response is attenuated in mutants with constitutive activation of the corticotropin-releasing factor receptor family homologue SEB-3. We show that activation of this receptor causes sex-common glutamatergic lumbar ganglion interneurons (LUA) to potentiate downstream male-specific reproduction circuits, allowing copulatory behaviours to partially override the light-induced escape responses in the male. SEB-3 activation in LUA also potentiates copulation during mild starvation. We suggest that SEB-3 activation allows C. elegans to acclimate to the environment and thus continue to execute innate behaviours even under non-optimal conditions.

σ1 receptor ligands control a switch between passive and active threat responses.

Humans and many animals show 'freezing' behavior in response to threatening stimuli. In humans, inappropriate threat responses are fundamental characteristics of several mental illnesses. To identify small molecules that modulate threat responses, we developed a high-throughput behavioral assay in zebrafish (Danio rerio) and evaluated 10,000 compounds for their effects on freezing behavior. We found three classes of compounds that switch the threat response from freezing to escape-like behavior. We then screened these for binding activity across 45 candidate targets. Using target profile clustering, we identified the sigma-1 (σ1) receptor as having a role in the mechanism of behavioral switching and confirmed that known σ1 ligands also disrupt freezing behavior. Furthermore, mutation of the gene encoding σ1 prevented the behavioral effect of escape-inducing compounds. One compound, which we call finazine, potently bound mammalian σ1 and altered threat-response behavior in mice. Thus, pharmacological and genetic interrogation of the freezing response revealed σ1 as a mediator of threat responses in vertebrates.

Long-term UVA eye irradiation causes decreased learning ability in mice.

BACKGROUND: Long-term eye radiation with ultraviolet A (UVA) denatures the cells of the cerebral hippocampus. However, the influence on memory and learning ability in mice is not known. METHODS: HR-1 mice were used. We irradiated the eyes or dorsal skin of the mice with UVA at a dose of 110 kJ/m(2) using an FL20SBLB-A lamp for 6, 12, and 24 months. RESULTS: The mean escape latency in the water maze was significantly increased in the UVA-irradiated mice in comparison with that seen in the controls. In the mice in which UVA eye irradiation was performed for 24 months, the depression of memory and learning ability was remarkable. The acetylcholinesterase activity, choline acetyltransferase activity, and acetylcholine content in the brain in the UVA eye-irradiated mice were significantly less than those observed in the control mice. Furthermore, during UVA eye irradiation, the levels of ß-amyloid (Aß), γ-secretase, which produces Aß peptide, and advanced glycation end products increased. Moreover, the effects of UVA eye irradiation increased with the duration of irradiation (or aging), and the introduction of glucose into the brain also decreased with UVA eye irradiation. CONCLUSIONS: These results indicate that UVA eye irradiation induces a decreased memory and learning ability in mice.

Differential peptide labeling (iTRAQ) in LC-MS/MS based proteomics in Daphnia reveal mechanisms of an antipredator response.

Daphnia, an important model organism for studies on ecology and evolution, has become a textbook example for inducible defenses against predators. Inducible defenses are widespread in nature, and the underlying molecular mechanisms for this plasticity in general and in particular in Daphnia are not fully understood. Here, we provide for the first time a combination of established life-history changes (LHC), which are induced by chemical cues of a predator (fish kairomones), in Daphnia with differential peptide labeling (iTRAQ) in LC-MS/MS based proteomics. The aim of the present study is the elucidation of proteins involved in specific antipredator responses in a predator-prey system of ecological relevance by high-throughput proteomics. To obtain a highly specific antifish response of Daphnia, highly purified fish kairomones were applied in the presence or absence of light. We were able to identify a set of functional proteins, which are likely to explain the kairomone-mediated and light-dependent LHC in Daphnia.

Relations between conspecific density and effects of ultraviolet-b radiation on tadpole size in the striped marsh frog.

Global increases in ultraviolet-B radiation (UVBR) associated with stratospheric ozone depletion are potentially contributing to the decline of numerous amphibian species around the world. Exposure to UVBR alone reduces survival and induces a range of sublethal effects in embryonic and larval amphibians. When additional environmental stressors are present, UVBR can have compounding negative effects. Thus, examination of the effects of UVBR in the absence of other stressors may substantially underestimate its potential to affect amphibians in natural habitats. We examined the independent and interactive effects of increased UVBR and high conspecific density would have embryonic and larval striped marsh frogs (Limnodynastes peronii). We exposed individuals to a factorial combination of low and high UVBR levels and low, medium, and high densities of striped marsh frog tadpoles. The response variables were time to hatching, hatching success, posthatch survival, burst-swimming performance of tadpoles (maximum instantaneous swim speed following an escape response), and size and morphology of tadpoles. Consistent with results of previous studies, we found that exposure to UVBR alone increased the time to hatching of embryos and reduced the burst-swimming performance and size of tadpoles. Similarly, increasing conspecific density increased the time to hatching of embryos and reduced the size of tadpoles, but had no effect on burst-swimming performance. The negative effect of UVBR on tadpole size was not apparent at high densities of tadpoles. This result suggests that tadpoles living at higher densities may invest relatively less energy in growth and thus have more energy to repair UVBR-induced damage. Lower densities of conspecifics increased the negative effects of UVBR on developing amphibians. Thus, low-density populations, which may include declining populations, may be particularly susceptible to the detrimental effects of increased UVBR and thus may be driven toward extinction faster than might be expected on the basis of results from single-factor studies.

Effect of sub-lethal exposure to ultraviolet radiation on the escape performance of Atlantic cod larvae (Gadus morhua).

UNLABELLED: The amount of ultraviolet (UV) radiation reaching the earth's surface has increased due to depletion of the ozone layer. Several studies have reported that UV radiation reduces survival of fish larvae. However, indirect and sub-lethal impacts of UV radiation on fish behavior have been given little consideration. We observed the escape performance of larval cod (24 dph, SL: 7.6±0.2 mm; 29 dph, SL: 8.2±0.3 mm) that had been exposed to sub-lethal levels of UV radiation vs. unexposed controls. Two predators were used (in separate experiments): two-spotted goby (Gobiusculus flavescens; a suction predator) and lion's mane jellyfish (Cyanea capillata; a "passive" ambush predator). Ten cod larvae were observed in the presence of a predator for 20 minutes using a digital video camera. Trials were replicated 4 times for goby and 5 times for jellyfish. Escape rate (total number of escapes/total number of attacks ×100), escape distance and the number of larvae remaining at the end of the experiment were measured. In the experiment with gobies, in the UV-treated larvae, both escape rate and escape distance (36%, 38±7.5 mm respectively) were significantly lower than those of control larvae (75%, 69±4.7 mm respectively). There was a significant difference in survival as well (UV: 35%, CONTROL: 63%). No apparent escape response was observed, and survival rate was not significantly different, between treatments (UV: 66%, CONTROL: 74%) in the experiment with jellyfish. We conclude that the effect and impact of exposure to sub-lethal levels of UV radiation on the escape performance of cod larvae depends on the type of predator. Our results also suggest that prediction of UV impacts on fish larvae based only on direct effects are underestimations.

[The influence of electromagnetic field on active avoidance reaction, biogenic amines and amino acids in brain of rats in spite of backround of food-stuff addition seratonus].

Amino acid preparation "Seratonus" not only regulates the action of stressors, but its intake in usual conditions strengthens an animals attitude toward danger, takes off development of excitement process, that is clearly shown by quantity and ratio of biogenic amines and amino acids. "Serotonus" develops optimal range of metabolism of above-mentioned substances that lead to right solve by animals of given tasks, in particular to improvement of active avoidance conditional reactions, that is clearly indicated by metabolism of amino acids as are phenylalanine and tryptophan, that is synthesizer of thirosine. While the latter takes part in synthesis of dopamine and noradrenalin and tryptophan regulates metabolism of serotonin. Hence these amines participate in improvement of memory processes.

Serotonergic mechanisms in the basolateral amygdala differentially regulate the conditioned and unconditioned fear organized in the periaqueductal gray.

The amygdala is an important filter for unconditioned and conditioned aversive information. The amygdala synthesizes the stimuli input from the environment and then signals the degree of threat that they represent to the dorsal periaqueductal gray (dPAG), which would be in charge of selecting, organizing and executing the appropriate defense reaction. In this study, we examined the influence of fluoxetine microinjections (1.75 and 3.5 nmol/0.2 microL) into the lateral (LaA) and basolateral (BLA) amygdaloid nuclei on the freezing and escape responses induced by electrical stimulation of the dPAG. Freezing behavior was also measured after the interruption of the electrical stimulation of the dPAG. On the following day, these rats were also submitted to a contextual fear paradigm to examine whether these microinjections would affect the conditioned freezing to contextual cues previously associated with foot shocks. Fluoxetine injections into both amygdaloid nuclei did not change the freezing and escape thresholds, but disrupted the dPAG-post-stimulation freezing. Moreover, the conditioned freezing was enhanced by fluoxetine. Whereas 5-HT mechanisms in the amygdala facilitate the acquisition of conditioned fear they inhibit the dPAG-post-stimulation freezing. However, the unconditioned fear triggered by activation of the dPAG is produced downstream of the amygdala. These findings have important implications for the understanding of the neurochemical substrates that underlie panic and generalized anxiety disorders.

A possible involvement of beta-endorphin, substance P, and serotonin in rat analgesia induced by extremely low frequency magnetic field.

Most of the research concerning magnetic antinociception was focused on brief exposure less than 1 h. The main purpose of the present study was to determine the effect of extremely low frequency (ELF) magnetic field (MF) repeated exposures on rats in inducing antinociception and to find the effective analgesic "time window." Meanwhile this investigation was to examine the role of central beta-endorphin, substance P, and 5-HT in magnetic analgesia. We found tail flick latencies (TFLs) increased significantly after the rats were exposed to 55.6 Hz, 8.1 mT magnetic field for 4 days, 6 h each day. The analgesic effects seemed to decrease gradually when the rats were exposed daily for another 10 days. Their levels of TFLs decreased within 1 day when the rats were removed after a 4-day exposure. The concentrations of hypothalamus beta-endorphin, substance P, and brainstem serotonin (5-HT) were increased significantly on Day 4. However, no differences were found when rats were exposed for another 10 days, and there were no significant increases when rats were removed after the fourth day of exposure and tested for nociception on Days 5 and 7 with no changes in the biochemical markers at 7 days. These results suggest that the ELF magnetic field has analgesic effect, but only on Days 3 and 4. The effect may be associated with increases in endogenous beta-endorphin, substance P, and 5-HT stimulated by the 55.6 Hz, 8.1 mT magnetic field.

Prior electrical stimulation of dorsal periaqueductal grey matter or deep layers of the superior colliculus sensitizes rats to anxiety-like behaviors in the elevated T-maze test.

Electrical stimulation of the dorsal periaqueductal grey matter (DPAG) and deep layers of the superior colliculus (DLSC) of the rat elicits anxiety-like reactions such as freezing and flight. The temporal course of the effects of the aversive electrical stimulation of the DPAG (5, 15 and 30 min afterward) and DLSC (5, 10 and 15 min afterward) on the defensive response of rats exposed to elevated T-maze were determined. The elevated T-maze generates two defensive behaviors, inhibitory avoidance and one-way escape, which have been related, respectively, to generalized anxiety and panic disorders. Prior electrical stimulation of the DPAG (15 min) and DLSC (5 min) enhanced inhibitory avoidance when compared to no-operated and sham animals, although not affecting escape. Therefore, stimulation of the DPAG and DLSC causes a heightened responsivity to anxiogenic stimulus, but not to panicogenic stimulus, inherent to elevated T-maze. These findings support the participation of the DPAG and DLSC in the elaboration of adaptive responses to stressful situations. Besides, the data supports the view that prior electrical stimulation of DPAG and DLSC is selective in sensitizing rats to anxiety-like behaviors, but not to panic-like behaviors in the elevated T-maze test.

Functional specializations within the tectum defense systems of the rat.

Here we review the differential contribution of the periaqueductal gray matter (PAG) and superior colliculus (SC) to the generation of rat defensive behaviors. The results of studies involving sine-wave and rectangular pulse electrical stimulation and chemical (NMDA) stimulation are summarized. Stimulation of SC and PAG produced freezing and flight behaviors along with exophthalmus (fully opened bulged eyes), micturition and defecation. The columnar organization of the PAG was evident in the results obtained. Defecation was elicited primarily by lateral PAG stimulation, while the remaining defensive behaviors were similarly elicited by lateral and dorsolateral PAG stimulation, although with the lowest thresholds in the dorsolateral column. Conversely, the ventrolateral PAG did not appear to participate in unconditioned defensive behaviors, which were only elicited by high intensity stimulation likely to encroach on adjacent regions. In the SC, the most important differences relative to the PAG were the lack of stimulation-evoked jumping in both intermediate and deep layers, and of NMDA-evoked galloping in intermediate layers. Therefore, we conclude that the SC may be only involved in the increased attentiveness (exophthalmus, immobility) and restlessness (trotting) of prey species exposed to the cues of a nearby predator. These responses may be distinct from the full-blown flight reaction that is mediated by the dorsolateral and lateral PAG. However, other evidences suggest the possible influences of stimulation schedule, environment dimensions and rat strain in determining outcomes. Overall our results suggest a dynamically organized representation of defensive behaviors in the midbrain tectum.

Disruption of left-right reciprocal coupling in the spinal cord of larval lamprey abolishes brain-initiated locomotor activity.

In this study, contributions of left-right reciprocal coupling mediated by commissural interneurons in spinal locomotor networks to rhythmogenesis were examined in larval lamprey that had longitudinal midline lesions in the rostral spinal cord [8 --> 30% body length (BL), relative distance from the head] or caudal spinal cord (30 --> 50% BL). Motor activity was initiated from brain locomotor command systems in whole animals or in vitro brain/spinal cord preparations. After midline lesions in the caudal spinal cord in whole animals and in vitro preparations, left-right alternating burst activity could be initiated in rostral and usually caudal regions of spinal motor networks. In in vitro preparations, blocking synaptic transmission in the rostral cord abolished burst activity in caudal hemi-spinal cords. After midline lesions in the rostral spinal cord in whole animals, left-right alternating muscle burst activity was present in the caudal and sometimes the rostral body. After spinal cord transections at 30% BL, rhythmic burst activity usually was no longer generated by rostral hemi-spinal cords. For in vitro preparations, very slow burst activity was sometimes present in isolated right and left rostral hemi-spinal cords, but the rhythmicity for this activity appeared to originate from the brain, and the parameters of the activity were significantly different from those for normal locomotor activity. In summary, in larval lamprey under these experimental conditions, left and right hemi-spinal cords did not generate rhythmic locomotor activity in response to descending inputs from the brain, suggesting that left-right reciprocal coupling contributes to both phase control and rhythmogenesis.

Long-term synaptic facilitation in defensive behavior command neurons in the snail during acquisition of sensitization depends on RNA synthesis.

Experiments on defensive behavior command neuron LP11 in semi-intact common snails preparations were performed to study transcription processes underlying the mechanisms of neuronal plasticity during acquisition of nociceptive sensitization. Application of nociceptive stimuli to the heads of control snails led to increases in neuron membrane excitability, along with biphasic changes in responses to tactile and chemical sensory stimulation--with depressed responses at the short-term stage and facilitation in the long-term stage of sensitization. Acquisition of sensitization after application of the RNA synthesis inhibitor actinomycin D (20 microM) to neuron LP11 led to complete suppression of synaptic facilitation in responses to sensory stimulation in the long-term stage of sensitization, but had no effect on the short-term depression of responses. However, the inhibitor had no effect on changes in membrane excitability in these conditions. Application of actinomycin D 30 min after the end of the sensitization acquisition process (1 h after the process started) had no effect on membrane excitability or synaptic facilitation. It is suggested that the induction of long-term synaptic facilitation in neuron LP11 during acquisition of nociceptive sensitization is dependent on RNA synthesis occurring in a narrow time interval, while changes in membrane excitability during the 2-3 h after acquisition of sensitization and short-term synaptic plasticity are independent of transcription processes.

Anterior cingulate cortical neuronal activity during perception of noxious thermal stimuli in monkeys.

It has been reported that the anterior cingulate cortex (ACC) has a variety of functions relating to pain as well as pain perception. However, the underlying mechanisms for those functions remain unclear. To elucidate the functional role of the ACC in pain perception and pain-related functions such as attention to pain and escape from pain, single neuronal activity was recorded from the ACC, and the behavioral correlates of this neuronal activity was studied. A total of 667 neurons were recorded from the ACC in awake behaving monkeys. Twenty-one had modulated activity during a heat-detection task. Eighteen of these increased their firing frequency following an increase in stimulus temperature, whereas three of them had decreased firing during heating of the face. Seventy-five percent of heat-evoked responses of heat-responsive ACC neurons were significantly depressed when monkeys detected the change in magnitude of illumination of a light presented on the front panel. The neuronal activity was significantly higher when monkeys escaped from a noxious heat stimulus than when the monkeys detected a small change in temperature (T2) above a larger initial shift (T1). No relationship between firing frequency and detection latency of the T2 stimulation was observed. These findings suggest that ACC nociceptive neurons are involved in attention to pain and escape from pain but not in the sensory discriminative aspect of pain.

Effect of number of tailshocks on learned helplessness and activation of serotonergic and noradrenergic neurons in the rat.

Adult male albino rats were exposed to varying numbers of tailshocks (0, 10, 50 or 100). The following day, their escape latencies in a shuttlebox were measured in order to estimate the degree of learned helplessness (LH) produced by the varying number of shocks. Only the groups exposed to 50 or 100 shocks displayed evidence of LH. In a parallel experiment, c-fos activation was used to determine the degree of activation of raphe serotonergic neurons (FosIR+5-HT) and locus coeruleus (LC) noradrenergic neurons (FosIR+TH) produced by the same shock conditions. Compared to unhandled cage controls, all shock groups (0 shocks was a restrained group) significantly activated both raphe and LC neurons. The 50 and 100 shock groups had significantly higher degrees of activation of serotonergic neurons in the rostral raphe groups and the LC than the 0 and 10 shock groups. These data are consistent with the hypothesis that activation of rostral raphe serotonergic neurons and LC noradrenergic neurons beyond a certain threshold may be critical for the development of LH. The relevance of these results for elucidating the neural bases of psychopathology is discussed.

[Analysis of the effects of low-intensity intermittent and continuous electromagnetic fields on behavioral acts in the mice].

The fact of modulating influence of low-intensity electromagnetic field on the behavioral acts in mice was established. The dependence of mice behavior on the modulating frequency, electromagnetic field parameters (intermittent or continuous) and on the time of adaplive reaction was found.