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RationaleThe phenylalkylamine hallucinogen (-)-2,5-dimethoxy-4-methylamphetamine (DOM) exhibits an inverted U-shaped dose-response curve for both head twitch response (HTR) and locomotor activity in mice. Accumulated studies suggest that HTR and locomotor hyperactivity induced by DOM are mainly caused by the activation of serotonin 5-hydroxytryptamine 2 A receptor (5-HT2A receptor). However, the mechanisms underlying the biphasic dose response of HTR and locomotor activity induced by DOM, particularly at high doses, remain unclear.ObjectivesThe primary objective of this study is to investigate the modulation of 5-HT2A/2C/1A receptors in HTR and locomotor activity, while also exploring the potential receptor mechanisms underlying the biphasic dose response of DOM.MethodsIn this study, we employed pharmacological methods to identify the specific 5-HT receptor subtypes responsible for mediating the biphasic dose-response effects of DOM on HTR and locomotor activity in C57BL/6J mice.ResultsThe 5-HT2A receptor selective antagonist (R)-[2,3-di(methoxy)phenyl]-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (M100907) (500 µg/kg, i.p.) fully blocked the HTR at every dose of DOM (0.615–10 mg/kg, i.p.) in C57BL/6J mice. M100907 (50 µg/kg, i.p.) decreased the locomotor hyperactivity induced by a low dose of DOM (0.625, 1.25 mg/kg, i.p.), but had no effect on the locomotor hypoactivity induced by a high dose of DOM (10 mg/kg) in C57BL/6J mice. The 5-HT2C antagonist 6-chloro-5-methyl-1-[(2-[2-methylpyrid-3yloxy]pyrid-5yl)carbamoyl]indoline (SB242084) (0.3, 1 mg/kg, i.p.) reduced the HTR induced by a dose of 2.5 mg/kg DOM, but did not affect the response to other doses. SB242084 (1 mg/kg, i.p.) significantly increased the locomotor activity induced by DOM (0.615–10 mg/kg, i.p.) in mice. The 5-HT1A antagonist N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]N-(2-pyridinyl) cyclohexane carboxamide maleate (WAY100635) (1 mg/kg, i.p.) increased both HTR and locomotor activity induced by DOM in mice. The 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) (1 mg/kg, i.p.) significantly reduced both the HTR and locomotor activity induced by DOM in mice. Additionally, pretreatment with the Gαi/o inhibitor PTX (0.25 µg/mouse, i.c.v.) enhanced the HTR induced by DOM and attenuated the effect of DOM on locomotor activity in mice.ConclusionsReceptor subtypes 5-HT2C and 5-HT1A are implicated in the inverted U-shaped dose-response curves of HTR and locomotor activity induced by DOM in mice. The biphasic dose-response function of HTR and locomotor activity induced by DOM has different mechanisms in mice.

Historically, most insect chronoecological research has used direct observations, cameras or infrared beam‐based monitors to quantify movement across timed intervals. Although some alternative DIY systems are cheaper than the current standard locomotor activity monitor, these options remain complicated to build and/or computationally intensive. We developed the imaging Locomotor Activity Monitor (iLAM), an affordable (~ $75 USD/unit) system for activity quantification. The iLAM utilizes a Raspberry Pi Zero W computer and night‐vision camera inside a flight cage to photograph a population of insects at user‐defined intervals. Open‐source, modular R‐scripts process the images and output a file containing the number, size, coordinate location and timestamp of all movements (blobs) identified between consecutive images. Output can be analysed directly or converted into the standard TriKinetics DAM format. We demonstrated the flexibility and power of the iLAM system by comparing diel and circadian activity of different insect species (fireflies: Photinus marginellus, P. greeni, P. obscurellus), ecotypes (moths: Ostrinia nubilalis) and sexes (moths: O. nubilalis). Data captured by only six iLAMs ($ 450) identified that peak activity of O. nubilalis females (AZT: 19.2 h) occurs significantly earlier than males (22.0 h). Additionally, male moths from a univoltine population exhibited a significantly shorter endogenous period length (AZT: 21.3 h) than males from a bivoltine genetic background (22.7 h). The iLAM will serve as a valuable tool for researchers seeking to measure locomotor activity across diverse species, sexes and populations in constant and changing environments.

Bergamot essential oil (BEO) is endowed with consistent and reproducible antinociceptive and anti-allodynic properties when administered via an inhalation route. However, the effects of its main constituents and of its decolored (DEC) and deterpenated (DET) fractions, which are enriched in limonene or in linalool and linalyl acetate, respectively, on spontaneous motor activity related to anxiety and on formalin-induced licking/biting biphasic behavior have never been investigated before. Therefore, the present research aims to characterize the role of BEO components on an experimental pain model that is relevant to clinical translation. Under our present experimental conditions, a paper filter disc soaked with different volumes of the phytocomplex and of its fractions that was applied at the edge of the observation chamber allowed the effects on the spontaneous motor activity and on the formalin-induced nocifensive response in ddY-strain mice to be studied. The present research demonstrated the effects of the DEC fraction of BEO on motor activity and on formalin-induced licking/biting behavior for the first time, proving that limonene is implicated in reduced motor activity and that it is important for the analgesic effect.

Excessive use of synthetic insecticides has resulted in environmental contamination and adverse effects on humans and other non-target organisms. Entomopathogenic fungi offer eco-friendly alternatives; however, their application for pest control requires significant advancement owing to limitations like slow killing time and effectiveness only when applied in higher amounts, whereas exposure to UV radiation, high temperature, and humidity can also reduce their viability and shelf-life. The nanoparticles synthesized using fungal extracellular extracts provide a new approach to use fungal pathogens. Our study focused on the synthesis of Metarhizium anisopliae –based silver nanoparticles (AgNPs) and evaluation of their efficiency on various physiological and behavioral parameters of the mosquito Aedes aegypti . The synthesis, size (27.6 d.nm, PDI = 0.209), zeta potential (− 24.3 mV), and shape of the AgNPs were determined through dynamic light scattering, scanning and transmission electron microscopic, and UV-visual spectroscopic analyses (432 nm). Our results showed significantly reduced survival (100% decrease in case of 3.2 and 1.8 μL/cm 2 volumes, and 60% decrease in case of 0.8 μL/cm 2 volume), phenoloxidase activity ( t = 39.91; p = 0.0001), and gut microbiota, with increased oxidative stress and cell apoptosis in AgNPs-challenged mosquitoes. Furthermore, the AgNPs-exposed mosquitoes presented a concentration-specific decrease in flight locomotor activity ( F = 17.312; p < 0.0001), whereas no significant changes in antifungal activity, self-grooming frequencies, or time spent were found. These findings enhance our understanding of mosquito responses to AgNPs exposure, and offer a more efficient mosquito control strategy using entomopathogenic fungi. Graphical Abstract

Abstract The aim of this study was to evaluate some physiological parameters as total locomotor activity and subcutaneous temperature in two different bovine breeds under tropical environment. In this study, total locomotor activity and subcutaneous temperature were evaluated in 18 clinically healthy female cows. All cows were divided into 3 groups: group A1 was constituted by 6 Holstein Bos taurus, group A2 was constituted by 6 female Herford Bos taurus cows, and group B was constituted by 6 female Bradford Bos indicus cows. Data recording of total locomotor activity (TLA) and subcutaneous temperature (ST) was performed by means of a subcutaneous thermometer (Thermocron) and an actimeter ACTIWATCH® (Cambridge Neurotechnology Ltd.) based on accelerometer technologies equipped on all subjects to record TLA. At the same time, thermal and hygrometric recording were carried out, considering the subtropical climate of Santa Fe. The application of GLM for statistical analysis showed a significant effect (P < 0.05) on statistical model and time of the day on TLA and ST for all groups; no significative effects on animal parameters were found for ST and TLA except for group B. Circadian parameters have been evaluated according to the single cosinor procedure of ST that showed a diurnal daily rhythmicity for all investigated groups and TLA which is focused almost during the photophase for groups A1 and A2 and during scotophase for group B. Considering different species and breeds, and different environmental conditions, this study suggested that some subjects may be much more able to adapt themselves to environmental stress than others. During their evolution from Bos taurus, zebu cattle (Bos indicus) have acquired genes that confer thermotolerance. Subjects from Bos indicus breeds are better able to regulate body temperature in response to heat stress than Bos taurus subjects.

The recent availability of commercial platforms for behavioral analyses in zebrafish larvae based on video-tracking technologies has exponentially increased the number of studies analyzing different behaviors in this model organism to assess neurotoxicity. Among the most commonly used assays in zebrafish larvae are basal locomotor activity (BLA) and visual motor responses (VMRs). However, the effect of different intrinsic and extrinsic factors that can significantly alter the outcome of these assays is still not well understood. In this work, we have analyzed the influence of age (5–8 days post-fertilization), time of day (8:00, 10:00, 12:00, 14:00; 16:00, 18:00, and 20:00 h), and experiment (three experiments performed at different days) on BLA and VMR results (4004 analyses for each behavior) in 143 larvae. The results from both behaviors were adjusted to a random-effects linear regression model using generalized least squares (GLSs), including in the model the effect of the three variables, the second-way interactions between them, and the three-way interaction. The results presented in this manuscript show a specific effect of all three intrinsic factors and their interactions on both behaviors, supporting the view that the most stable time period for performing these behavioral assays is from 10:00 am to 04:00 pm, with some differences depending on the age of the larva and the behavioral test.

Abstract Human activities involving noise emission can affect wild animals. European mink was exposed to road noise and human voice playbacks to analyze how sound intensity level and duration of both noises altered the time that individuals were active and if their fecal cortisol metabolite (FCM) levels varied. A Hierarchical Analysis Cluster was performed to establish 2 mink groups with respect to both noise source type: short duration/low intensity (SL) and long duration/high intensity (LH). We performed general linear mixed models to evaluate the variation in locomotor activity duration (s) and FCM (nanogram per gram) levels, respectively. The results showed both road noise and human voices decreased locomotor activity duration in SL more sharply compared with LH, and human voices were the triggers that induced the most pronounced response to both exposure conditions. FCM (ng/g) levels increased in SL compared with LH during road noise while the opposite happened during human voices. Differences based on sex and age of individuals were observed. In conclusion, noise characteristics given by the sound type determined the variations in locomotor activity duration while noise exposure level determined the variations in FCM (ng/g) levels. Attention should be paid to noisy activities (e.g., recreational activities for visitors in protected natural areas) and loud groups of people to conserve wildlife, especially noise sensitive species.

The mammalian circadian clock system regulates physiological function. Crude drugs, containing Polygalae Radix, and Kampō, combining multiple crude drugs, have been used to treat various diseases, but few studies have focussed on the circadian clock. We examine effective crude drugs, which cover at least one or two of Kampō, for the shortening effects on period length of clock gene expression rhythm, and reveal the mechanism of shortening effects. We prepared 40 crude drugs. In the in vitro experiments, we used mouse embryonic fibroblasts from PERIOD2::LUCIFERASE knock-in mice (background; C57BL/6J mice) to evaluate the effect of crude drugs on the period length of core clock gene, Per2, expression rhythm by chronic treatment (six days) with distilled water or crude drugs (100 μg/mL). In the in vivo experiments, we evaluated the free-running period length of C57BL/6J mice fed AIN-93M or AIN-93M supplemented with 1% crude drug (6 weeks) that shortened the period length of the PERIOD2::LUCIFERASE expression rhythm in the in vitro experiments. We found that Polygalae Radix (ED 50 : 24.01 μg/mL) had the most shortened PERIOD2::LUCIFERASE rhythm period length in 40 crude drugs and that the CaMKII pathway was involved in this effect. Moreover, long-term feeding with AIN-93M+Polygalae Radix slightly shortened the free-running period of the mouse locomotor activity rhythm. Our results indicate that Polygalae Radix may be regarded as a new therapy for circadian rhythm disorder and that the CaMKII pathway may be regarded as a target pathway for circadian rhythm disorders.

Background Mutants which carry mutations in genes encoding mitochondrial ligases MUL1 and PARKIN are convenient Drosophila models of Parkinson’s disease (PD). In several studies it has been shown that in Parkinson’s disease sleep disturbance occurs, which may be the result of a disturbed circadian clock. Results We found that the ROS level was higher, while the anti-oxidant enzyme SOD1 level was lower in mul1 A6 and park 1 mutants than in the white mutant used as a control. Moreover, mutations of both ligases affected circadian rhythms and the clock. The expression of clock genes per , tim and clock and the level of PER protein were changed in the mutants. Moreover, expression of ATG5, an autophagy protein also involved in circadian rhythm regulation, was decreased in the brain and in PDF-immunoreactive large ventral lateral clock neurons. The observed changes in the molecular clock resulted in a longer period of locomotor activity rhythm, increased total activity and shorter sleep at night. Finally, the lack of both ligases led to decreased longevity and climbing ability of the flies. Conclusions All of the changes observed in the brains of these Drosophila models of PD, in which mitochondrial ligases MUL1 and PARKIN do not function, may explain the mechanisms of some neurological and behavioural symptoms of PD.

The two hemispheres of the vertebrate brain are connected through several commissures. Although the anterior commissure (AC) is the most conserved white matter structure in the brains of different vertebrates, its complete physiological functionality remains elusive. Since the AC is involved in the connection between two amygdalae and because amygdalae are critical for emotional behaviors and social interaction, we assessed amygdalar activity and function to investigate the physiological role of the AC. We first performed electrophysiological recording on mouse brains to demonstrate that the AC delivers a positive signal to facilitate synaptic responses and to recruit basolateral amygdalar neurons via glutamatergic synapses. Transection was then undertaken to investigate the role of the AC . Results from optogenetic stimulation suggest that AC transection impairs mutual activation between two basolateral amygdalae. Behavioral analyses were then used to assess if AC surgical lesioning results in hyperactivity, anxiety, social reduction or learning/memory impairment, which are behavioral features associated with neuropsychiatric disorders, such as autism spectrum disorders. We found that AC transection results in higher locomotor activity, aberrant social interaction and reduced associative memory, but not anxiety. Moreover, systemic administration of D-cycloserine, a coagonist of N-methyl-D-aspartate receptor, ameliorated auditory fear memory in AC-transected mice, reinforcing our evidence that the AC potentiates the activity of basolateral amygdalae. Our study suggests that the AC regulates basolateral amygdalar activity and influences neuropsychiatry-related behaviors in mice.