Behavioral Responses In Mice Research Articles

Connectomics Investigation of Role of Parabrachial Nucleus Aversion using Optogenetics: A Experiment Proposal

In recent decades, there has been a notable increase in research activity focused on the aversive and reward circuits of the brain. The evidence indicates that the parabrachial nucleus (PBn) is a crucial element in the generation of aversion responses in mice. Following successful stimulation of the PBn, mice display increased aversive and stress-related ehaviours. The study employs Fos TRAP2-ChR2 mice, opioids such as oxycodone, and a range of imaging technologies to monitor and manipulate neural pathways. Behavioral tests and optogenetic manipulation are used to assess real-time behavioral responses in mice, with the aim of uncovering the neural pathways and mechanisms involved in PBn-mediated aversion. Nevertheless, the underlying mechanisms of these behaviours remain unclear. The objective of this study is to investigate the connectomics of the parabrachial nucleus (PBn) in order to determine its association with regions involved in motivation, such as the nucleus ambiguus or the amygdala. It is hypothesised that this association may lead to aversion by affecting these regions. An alternative hypothesis is that if the parabrachial nucleus is connected to areas that are not directly related to it, it may be considered a motivational area in its own right. The identification of these connections will elucidate the underlying mechanisms of aversion mediated by PBn, thereby providing a deeper understanding of the neural basis of aversive behaviour and its regulation.

Betaine improves METH-induced depressive-like behavior and cognitive impairment by alleviating neuroinflammation via NLRP3 inflammasome inhibition

Methamphetamine abuse has been associated with central nervous system damage, contributing to the development of neuropsychiatric disorders such as depressive-like behavior and cognitive impairment. With the escalating prevalence of METH abuse, there is a pressing need to explore effective therapeutic interventions. Thus, the objective of this research was to investigate whether betaine can protect against depressive-like behavior and cognitive impairment induced by METH. Following intraperitoneal injections of METH in mice, varying doses of betaine were administered. Subsequently, the behavioral responses of mice and the impact of betaine intervention on METH-induced neural damage, synaptic plasticity, microglial activation, and NLRP3 inflammatory pathway activation were assessed. Administration 30 mg/kg and 100 mg/kg of betaine ameliorated METH-induced depressive-like behaviors in the open field test, tail suspension test, forced swimming test, and sucrose preference test and cognitive impairment in the novel object recognition test and Barnes maze test. Moreover, betaine exerted protective effects against METH-induced neural damage and reversed the reduced synaptic plasticity, including the decline in dendritic spine density, as well as alterations in the expression of hippocampal PSD95 and Synapsin-1. Additionally, betaine treatment suppressed hippocampal microglial activation induced by METH. Likewise, it also inhibited the activation of the hippocampal NLRP3 inflammasome pathway and reduced IL-1β and TNF-α release. These results collectively suggest that betaine's significant role in mitigating depressive-like behavior and cognitive impairment resulting from METH abuse, presenting potential applications in the prevention and treatment of substance addiction.

The synthetic cathinones MDPHP and MDPV: Comparison of the acute effects in mice, in silico ADMET profiles and clinical reports

The 3,4-methylenedioxy-alpha-pyrrolidinohexanophenone (MDPHP) is a synthetic cathinone closely related to 3,4-methylenedioxypyrovalerone (MDPV), one of the most common synthetic cathinones present in the “bath salts”. MDPHP has recently gained attention due to increasing seizures and involvement in human intoxications which occurred in Europe and Italy in the last years, but currently there is a lack of information about its pharmaco-toxicological effects. With the aim at filling this gap, the present study is endeavoured to (i) evaluate the effects of acute administration of MDPHP (0.01–20 mg/kg; i.p.) on behaviour, cardiorespiratory and cardiovascular parameters in CD-1 male mice, comparing them to those observed after administration of MDPV; (ii) predict the ADMET profile of the two analogues using the Plus ADMET Predictor®; (iii) present clinical data related to MDPHP and MDPV-induced intoxications recorded between 2011 and 2023 by the Pavia Poison Control Centre (PCC) − National Toxicology Information Centre (Istituti Clinici Scientifici Maugeri, IRCCS Pavia, Italy). Our results substantiated that MDPHP and MDPV similarly affect sensorimotor and behavioural responses in mice, importantly increased locomotion and induced aggressive behaviour, and, at higher dosage, increased heart rate and blood pressure. These findings are in line with those observed in humans, revealing severe toxidromes typically characterized by Central Nervous System (CNS) alterations (behavioural/neuropsychiatric symptoms), including psychomotor agitation and aggressiveness, cardiovascular and respiratory disorders (e.g. tachycardia, hypertension, dyspnoea), and other peripheral symptoms (e.g. hyperthermia, acidosis, rhabdomyolysis).

Mutation of novel ethanol-responsive lncRNA Gm41261 impacts ethanol-related behavioral responses in mice.

Chronic alcohol exposure results in widespread dysregulation of gene expression that contributes to the pathogenesis of Alcohol Use Disorder (AUD). Long noncoding RNAs are key regulators of the transcriptome that we hypothesize coordinate alcohol-induced transcriptome dysregulation and contribute to AUD. Based on RNA-Sequencing data of human prefrontal cortex, basolateral amygdala and nucleus accumbens of AUD versus non-AUD brain, the human LINC01265 and its predicted murine homolog Gm41261 (i.e., TX2) were selected for functional interrogation. We tested the hypothesis that TX2 contributes to ethanol drinking and behavioral responses to ethanol. CRISPR/Cas9 mutagenesis was used to create a TX2 mutant mouse line in which 306 base-pairs were deleted from the locus. RNA analysis revealed that an abnormal TX2 transcript was produced at an unchanged level in mutant animals. Behaviorally, mutant mice had reduced ethanol, gaboxadol and zolpidem-induced loss of the righting response and reduced tolerance to ethanol in both sexes. In addition, a male-specific reduction in two-bottle choice every-other-day ethanol drinking was observed. Male TX2 mutants exhibited evidence of enhanced GABA release and altered GABAA receptor subunit composition in neurons of the nucleus accumbens shell. In C57BL6/J mice, TX2 within the cortex was cytoplasmic and largely present in Rbfox3+ neurons and IBA1+ microglia, but not in Olig2+ oligodendrocytes or in the majority of GFAP+ astrocytes. These data support the hypothesis that TX2 mutagenesis and dysregulation impacts ethanol drinking behavior and ethanol-induced behavioral responses in mice, likely through alterations in the GABAergic system.

Cryopreserved Placental Derivatives Improve Quality of Life and Behavioral Responses in Mice with Chemotherapy-Induced Ovarian Failure

Cryopreserved Placental Derivatives Improve Quality of Life and Behavioral Responses in Mice with Chemotherapy-Induced Ovarian Failure

Degrading stimuli by reducing image resolution impairs performance in a rodent continuous performance test

Attention is a cognitive domain often disrupted in neuropsychiatric disorders and continuous performance tests (CPTs) are common clinical assays of attention. In CPTs, participants produce a behavioral response to target stimuli and refrain from responding to non-target stimuli. Performance in CPTs is measured as the ability to discriminate between targets and non-targets. Rodent versions of CPTs (rCPTs) have been validated with both anatomical and pharmacological studies, providing a translational platform for understanding attention function. In humans, stimulus degradation, the inclusion of visual noise in the image to reduce resolution, in CPTs impairs performance. Reduced image contrast, changes in the relative luminescence of elements in the image, has been used in rCPTs to test similar constructs, but, to our knowledge, reduced image resolution has not been tested in an rCPT. In this study, we tested multiple levels of stimulus degradation in a touchscreen version of the rCPT in mice. We found that stimulus degradation significantly decreased performance in males and females. Specifically, we found decreased stimulus discrimination and increases in hit reaction time and reaction time variability. These findings are in line with the effects of stimulus degradation in human studies. These data extend the utility and translational value of the family of rCPTs by demonstrating that stimulus degradation in the form of reduced image resolution produces qualitatively similar behavioral responses in mice as those in previous human studies.

Hemoglobin in the blood acts as a chemosensory signal via the mouse vomeronasal system

The vomeronasal system plays an essential role in sensing various environmental chemical cues. Here we show that mice exposed to blood and, consequently, hemoglobin results in the activation of vomeronasal sensory neurons expressing a specific vomeronasal G protein-coupled receptor, Vmn2r88, which is mediated by the interaction site, Gly17, on hemoglobin. The hemoglobin signal reaches the medial amygdala (MeA) in both male and female mice. However, it activates the dorsal part of ventromedial hypothalamus (VMHd) only in lactating female mice. As a result, in lactating mothers, hemoglobin enhances digging and rearing behavior. Manipulation of steroidogenic factor 1 (SF1)-expressing neurons in the VMHd is sufficient to induce the hemoglobin-mediated behaviors. Our results suggest that the oxygen-carrier hemoglobin plays a role as a chemosensory signal, eliciting behavioral responses in mice in a state-dependent fashion.

Perinatal Food Deprivation Modifies the Caloric Restriction Response in Adult Mice Through Sirt1

Variations in the availability of nutritional resources in animals can trigger reversible adjustments, which in the short term are manifested as behavioral and physiological changes. Several of these responses are mediated by Sirt1, which acts as an energy status sensor governing a global genetic program to cope with changes in nutritional status. Growing evidence suggests a key role of the response of the perinatal environment to caloric restriction in the setup of physiological responses in adulthood. The existence of adaptive predictive responses has been proposed, which suggests that early nutrition could establish metabolic capacities suitable for future food-scarce environments. We evaluated how perinatal food deprivation and maternal gestational weight gain impact the transcriptional, physiological, and behavioral responses in mice, when acclimated to caloric restriction in adulthood. Our results show a strong predictive capacity of maternal weight and gestational weight gain, in the expression of Sirt1 and its downstream targets in the brain and liver, mitochondrial enzymatic activity in skeletal muscle, and exploratory behavior in offspring. We also observed differential responses of both lactation and gestational food restriction on gene expression, thermogenesis, organ masses, and behavior, in response to adult caloric restriction. We conclude that the early nutritional state could determine the magnitude of responses to food scarcity later in adulthood, mediated by the pivotal metabolic sensor Sirt1. Our results suggest that maternal gestational weight gain could be an important life history trait and could be used to predict features that improve the invasive capacity or adjustment to seasonal food scarcity of the offspring.

The effects of combined administration of human umbilical cord-derived multipotent mesenchymal stromal cells and melatonin or fibroblast growth factor-2 to aged mice with a toxic cuprizone model of demyelination

The effect of transplantation of umbilical cord-derived multipotent mesenchymal stromal cells (UC-MMSCs) to patients with demyelinating diseases depends on the age of the recipient and can change under the influence of hormones or growth factors. Purpose. To investigate the effect of exogenous melatonin and recombinant human fibroblast growth factor-2 (rhFGF-2) on the effects of UC-MMSCs transplanted into aged mice with an experimental model of multiple sclerosis. Material and methods. 129/Sv mice, 15-17 months old, received the neurotoxin cuprizone with food for 3 weeks. From the 10th day of the cuprizone diet, 5•105 UC-MMSCs were injected intravenously. From the 11th day they received melatonin at 600 p.m. or rhFGF-2. The behavioral parameters were evaluated in the open field test and rotarod test. In the brain, the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and the level of malondialdehyde (MDA) were assessed. Results. Cuprizone intake reduces the behavioral response in mice compared to the intact group. The transplantation of UC-MMSCs increases the number of rearings and muscle tone in mice. Melatonin injections enhance the effects of cells on these parameters, as well as increase the motor and emotional activity of animals. The injection of rhFGF-2 preserves the effect of cells on behavioral response and increases locomotor activity in mice. After the injection of UC-MMSCs with melatonin or rhFGF-2, the content of MDA in the brain decreases and the activity of antioxidant enzymes increases, this is more significant under the influence of melatonin. Conclusion. Exogenous melatonin and rhFGF-2 improve the effects of transplanted UC-MMSCs on behavioral responses and brain antioxidant defenses in aged mice with cuprizone diet. At the same time, the positive effect of the combination of cells with melatonin is more pronounced.

Anxiety-related defensive behavioral responses in mice selectively bred for High and Low Activity.

High and Low Activity strains of mice (displaying low and high anxiety-like behavior, respectively) with 7.8-20 fold differences in open-field activity were selected and subsequently inbred to use as a genetic model for studying anxiety-like behavior in mice (DeFries et al., 1978, Behavior Genetics, 8:3-13). These strains exhibited differences in other anxiety-related behaviors as assessed using the light-dark box, elevated plus-maze, mirror chamber, and elevated square-maze tests (Henderson et al., 2004, Behavior Genetics, 34: 267-293). The purpose of these experiments was three-fold. First, we repeated a 6-day behavioral battery using updated equipment and software to confirm the extreme differences in anxiety-like behaviors. Second, we tested novel object exploration, a measure of anxiety-like behavior that does not rely heavily on locomotion. Third, we conducted a home cage wheel running experiment to determine whether these strains differ in locomotor activity in a familiar, home cage environment. Our behavioral test battery confirmed extreme differences in multiple measures of anxiety-like behaviors. Furthermore, the novel object test demonstrated that the High Activity mice exhibited decreased anxiety-like behaviors (increased nose pokes) compared to Low Activity mice. Finally, male Low Activity mice ran nearly twice as far each day on running wheels compared to High Activity mice, while female High and Low Activity mice did not differ in wheel running. These results support the idea that the behavioral differences between High and Low Activity mice are likely to be due to anxiety-related factors and not simply generalized differences in locomotor activity.

Complex effect of caffeine and dioxidine on behavioral responses in mice in Porsolt test

Relevance. In light of the popularization of the use of caffeine-containing products, the question of the combined use of caffeine with substances exhibiting a toxic effect remains open. The doses of caffeine, which have a pronounced antidepressant effect, are also insufficiently studied. The aim of the study was to study the effect of repeated administration of caffeine and dioxidine on the behavioral responses of mice in the Porsolt test. Materials and methods. The experiment was carried out on 36 outbred male mice, divided into 6 groups. Experimental groups for 15 days of the study received caffeine at a dose of 40 mg/kg (first) or 100 mg/kg (second), dioxidine at a dose of 200 mg/kg (third), together with caffeine 40 mg/kg or 100 mg/kg, and dioxidine (fourth and fifth groups, respectively). The animals of the control group were injected with saline. To study the behavior, the Porsolt test was carried out, evaluating the following indicators on the 1st, 8th and 15th days of the experiment: the total time of immobilization, active swimming, climb, the number of grooming and shaking off acts. Results . The administration of caffeine at a dose of 40 mg/kg caused an increase in the time of active swimming and a decrease in the duration of immobilization on the 8th and 15th days. When caffeine was used at a dose of 100 mg/kg, an increase in the time of active swimming was noted with a single exposure, with an experiment duration of 8-15 days, an increase in the duration of immobilization was observed. Dioxidine caused a significant decrease in the time of active swimming and an increase in the duration of immobilization during all days of the experiment. The combined use of caffeine (40 mg/kg and 100 mg/kg) and dioxidine on the 1st day led to a decrease in immobilization and the time of active swimming. In both groups, 100 % animal mortality was observed by the 15th day. Conclusion. The results of the study indicate the presence of an antidepressant effect in caffeine at a dose of 40 mg/kg on the 8th and 15th days of the experiment and the absence of this effect in caffeine at a dose of 100 mg/kg with a duration of administration of 8-15 days. The use of dioxidine led to the absence of antidepressant activity and the presence of the opposite effect. The combined administration of caffeine (40 mg/kg and 100 mg/kg) and dioxidine led to 100 % mortality in the experimental groups by the 15th day of the experiment

Negative feedback control of neuronal activity by microglia.

Microglia, the brain’s resident macrophages, help to regulate brain function by removing dying neurons, pruning non-functional synapses, and producing ligands that support neuronal survival1. Here we show that microglia are also critical modulators of neuronal activity and associated behavioural responses in mice. Microglia respond to neuronal activation by suppressing neuronal activity, and ablation of microglia amplifies and synchronizes the activity of neurons, leading to seizures. Suppression of neuronal activation by microglia occurs in a highly region-specific fashion and depends on the ability of microglia to sense and catabolize extracellular ATP, which is released upon neuronal activation by neurons and astrocytes. ATP triggers the recruitment of microglial protrusions and is converted by the microglial ATP/ADP hydrolysing ectoenzyme CD39 into AMP; AMP is then converted into adenosine by CD73, which is expressed on microglia as well as other brain cells. Microglial sensing of ATP, the ensuing microglia-dependent production of adenosine, and the adenosine-mediated suppression of neuronal responses via the adenosine receptor A1R are essential for the regulation of neuronal activity and animal behaviour. Our findings suggest that this microglia-driven negative feedback mechanism operates similarly to inhibitory neurons and is essential for protecting the brain from excessive activation in health and disease.

Physiological and Behavioral Responses to Vocalization Playback in Mice.

In mice, the caller’s production of social vocalizations has been extensively studied but the effect of these vocalizations on the listener is less understood, with playback studies to date utilizing one vocalization category or listeners of one sex. This study examines how several categories of mouse vocalizations affect listeners of both sexes to better understand the communicative functions of these vocal categories. We examined physiological and behavioral responses of male and female CBA/CaJ mice to playback of four social vocalization categories: ultrasonic vocalizations (USVs), low-frequency harmonic calls, mid-frequency vocalizations, and noisy calls. Based on the conditions under which these calls are emitted, we hypothesized that playback of these vocal categories would have differential effects on the listeners. In females, playback of all four vocalization categories increased stress hormone levels (corticosterone), but only the non-USV categories increased corticosterone in males. The magnitude of corticosterone increase in non-USV trials was greater in females than in males. In open field tests, all four vocal categories decreased central ambulation in males and females, indicating an increase in anxiety-related behavior. Further, we found that the proportions of USVs emitted by subjects, but not their overall calling rates, were affected by playback of some vocal categories, suggesting that vocalization categories have different communication content. These results show that, even in the absence of behavioral and acoustic contextual features, each vocal category evokes physiological and behavioral responses in mice, with some differences in responses as a function of the listener’s sex and playback signal. These findings suggest that at least some of the vocal categories have distinct communicative functions.

Keratinocytes contribute to normal cold and heat sensation.

Keratinocytes are the most abundant cell type in the epidermis, the most superficial layer of skin. Historically, epidermal-innervating sensory neurons were thought to be the exclusive detectors and transmitters of environmental stimuli. However, recent work from our lab (Moehring et al., 2018) and others (Baumbauer et al., 2015) has demonstrated that keratinocytes are also critical for normal mechanotransduction and mechanically-evoked behavioral responses in mice. Here, we asked whether keratinocyte activity is also required for normal cold and heat sensation. Using calcium imaging, we determined that keratinocyte cold activity is conserved across mammalian species and requires the release of intracellular calcium through one or more unknown cold-sensitive proteins. Both epidermal cell optogenetic inhibition and interruption of ATP-P2X4 signaling reduced reflexive behavioral responses to cold and heat stimuli. Based on these data and our previous findings, keratinocyte purinergic signaling is a modality-conserved amplification system that is required for normal somatosensation in vivo.

The Effect of Quercetin on Metabolism and Behavioral Responses in Mice with Normal and Impaired Leptin Reception

We studied the effects of quercetin (Q) on the behavioral responses and integral and metabolic indices in male db/db mice genetically predisposed to obesity and C57Bl/6J mice fed a diet with an excessive energy value. We examined forepaw grip strength, exploratory activity, anxiety, and short-term memory, as well as integral and biochemical indices, and the contents of leptin, ghrelin, and IL-10. We found differences between two models of obesity in the integral and biochemical indices and behavioral responses to Q treatment. In the db/db mice, they were associated with knockout of the Lepr gene encoding the leptin receptor, which prevented the neurotropic and regulatory functions of leptin. These effects of Q detected should be considered when justifying the dose and timing of this nutraceutical in clinical practice.

Toll-like receptor 4 deficiency ameliorates β2-microglobulin induced age-related cognition decline due to neuroinflammation in mice

Toll-like receptor 4 (TLR4) is a crucial receptor in neuroinflammation and apoptotic neuronal death, and increasing evidences indicated that β2-microglobulin (B2M) is thought to be a major contributor to age-related cognitive decline. In present study, we designed to investigate the effects of TLR4 on B2M-induced age-related cognitive decline. Wild-type (WT) C57BL/6, TLR4 knockout (TLR4 -KO) mice and hippocampal neurons from the two type mice were respectively divided into two groups: (1) Veh group; (2) B2M-treated group. The behavioral responses of mice were measured using Morris Water Maze. Hippocampal neurogenesis and neuronal damage, inflammatory response, apoptosis, synaptic proteins and neurotrophic factors, and TLR4/MyD88/NF-κB signaling pathway proteins were examined using molecular biological or histopathological methods. The results showed that WT mice received B2M in the DG exhibited age-related cognitive declines, increased TLR4 mRNA expression and high levels of interleukin-1β (IL-1β), tumor necrosis factor-alpha (TNF-α) and apoptotic neuronal death in the hippocampus, which were partially attenuated in TLR4-KO mice. Moreover, in absence of TLR4, B2M treatment improved hippocampus neurogenesis and increased synaptic related proteins. Our cell experiments further demonstrated that deletion of TLR4 could significantly increase synaptic related protein, decrease neuroinflammatory fators, inhibited apoptotic neuronal death, and regulated MyD88/NF-κB signal pathway after B2M treatment. In summary, our results support the TLR4 contributes to B2M-induced age-related cognitive decline due to neuroinflammation and apoptosis through TLR4/MyD88/NF-κB signaling pathway via a modulation of hippocampal neurogenesis and synaptic function. This may provide an important neuroprotective mechanism for improving age-related cognitive decline.

Augmentation effect of ketamine by guanosine in the novelty-suppressed feeding test is dependent on mTOR signaling pathway

The ketamine's potential for the treatment of refractory depression and anxiety has been considered one the most important discoveries in the last years, however, repeated use of ketamine is limited due to its side/adverse effects. Therefore, the search for effective augmentation strategies that may reduce ketamine doses is welcome. Therefore, this study sought to augment the effect of ketamine by guanosine in the novelty-suppressed feeding (NSF) test, a behavioral paradigm able to detect depression/anxiety-related behavior. Acute administration of guanosine (0.05 mg/kg, p.o.), similar to ketamine (1 mg/kg, i.p.), produced a rapid behavioral response in mice submitted to NSF test. Moreover, the coadministration of sub-effective doses of guanosine (0.01 mg/kg, p.o.) and ketamine (0.1 mg/kg, i.p.) was effective in mice submitted to NSF test. Subsequently, the intracellular mechanism underpinning the augmentation effect of ketamine by guanosine was investigated. Our results suggest that augmentation response of ketamine by guanosine in the NSF test probably involves the activation of mTOR signaling, since the treatment with rapamycin (0.2 nmol/site, i.c.v., a selective mTOR inhibitor) completely abolished this effect. This augmentation strategy also increased mTOR phosphorylation (Ser2448) in the hippocampus, reinforcing the role of mTOR in this augmentation response. However, no changes in the p70S6K, PSD-95, GluA1, and synapsin immunocontents were found in the hippocampus of ketamine plus guanosine-treated mice. Overall, results provide evidence that guanosine is able to augment the effect of ketamine in the NSF test via mTOR activation, a finding that might have therapeutic implications for the management of depression/anxiety.

β1-adrenergic receptors mediate plasma acyl-ghrelin elevation and depressive-like behavior induced by chronic psychosocial stress.

The ghrelin system is a key component of the mood and metabolic responses to chronic psychosocial stress. For example, circulating acyl-ghrelin rises in several rodent and human stress models, administered acyl-ghrelin induces antidepressant-like behavioral responses in mice, and mice with deleted ghrelin receptors (GHSRs) exhibit exaggerated depressive-like behaviors, changed eating behaviors, and altered metabolism in response to chronic stress. However, the mechanisms mediating stress-induced rises in ghrelin are unknown and ghrelin's antidepressant-like efficacy in the setting of chronic stress is incompletely characterized. Here, we used a pharmacological approach in combination with a 10-day chronic social defeat stress (CSDS) model in male mice to investigate whether the sympathoadrenal system is involved in the ghrelin response to stress. We also examined the antidepressant-like efficacy of administered ghrelin and the synthetic GHSR agonist GHRP-2 during and/or after CSDS. We found that administration of the β1-adrenergic receptor (β1AR) blocker atenolol during CSDS blunts the elevation of plasma acyl-ghrelin and exaggerates depressive-like behavior. Neither acute injection of acyl-ghrelin directly following CSDS nor its chronic administration during or after CSDS nor chronic delivery of GHRP-2 during and after CSDS improved stress-induced depressive-like behavior. Thus, β1ARs drive the acyl-ghrelin response to CSDS, but supplementing the natural increases in acyl-ghrelin with exogenous acyl-ghrelin or GHSR agonist does not further enhance the antidepressant-like actions of the endogenous ghrelin system in the setting of CSDS.

Cbln2 and Cbln4 are expressed in distinct medial habenula-interpeduncular projections and contribute to different behavioral outputs

Cerebellins are important neurexin ligands that remain incompletely understood. Two critical questions in particular remain unanswered: do different cerebellins perform distinct functions, and do these functions act in the initial establishment of synapses or in rendering nascent synapses capable of normal synaptic transmission? Here we show that in mice, Cbln2 and Cbln4 are expressed in the medial habenula (MHb) nucleus in different types of neurons that project to distinct target neurons in the interpeduncular nucleus. Conditional genetic deletion of Cbln2 in the MHb impaired synaptic transmission at Cbln2+ synapses in the interpeduncular neurons within 3 wk, but decreased synapse numbers only after 3 mo, suggesting a functional, but not a structural, requirement for Cbln2 in synapses formed by Cbln2-expressing neurons. In contrast, genetic deletions of Cbln4 in the MHb had no major effect on synaptic transmission or synapse numbers in interpeduncular target neurons. Nevertheless, MHb ablation of both Cbln2 and Cbln4 significantly impaired behavioral responses in mice, but affected different types of behaviors. Specifically, Cbln2 MHb deletions decreased spatial learning, as measured in the water T-maze, whereas Cbln4 MHb deletions increased anxiety levels, as monitored in the open field test and elevated plus maze. Thus, Cbln2 and Cbln4 are expressed in distinct MHb neurons that contribute to different behaviors.

Transient Magnetothermal Neuronal Silencing Using the Chloride Channel Anoctamin 1 (TMEM16A).

Determining the role and necessity of specific neurons in a network calls for precisely timed, reversible removal of these neurons from the circuit via remotely triggered transient silencing. Previously, we have shown that alternating magnetic field mediated heating of magnetic nanoparticles, bound to neurons, expressing temperature-sensitive cation channels TRPV1 remotely activates these neurons, evoking behavioral responses in mice. Here, we demonstrate how to apply magnetic nanoparticle heating to silence target neurons. Rat hippocampal neuronal cultures were transfected to express the temperature gated chloride channel, anoctamin 1 (TMEM16A). Spontaneous firing was suppressed within seconds of alternating magnetic field application to anoctamin 1 (TMEM16A) channel expressing, magnetic nanoparticle decorated neurons. Five seconds of magnetic field application leads to 12 s of silencing, with a latency of 2 s and an average suppression ratio of more than 80%. Immediately following the silencing period spontaneous activity resumed. The method provides a promising avenue for tether free, remote, transient neuronal silencing in vivo for both scientific and therapeutic applications.