Freezing Response Research Articles

Context-induced renewal of passive but not active coping behaviours in the shock-probe defensive burying task.

Renewal is the return of extinguished responding after removal from the extinction context. Renewal has been extensively studied using classical aversive conditioning procedures that measure a passive freezing response to an aversive conditioned stimulus. However, coping responses to aversive stimuli are complex and can be reflected in passive and active behaviours. Using the shock-probe defensive burying task, we investigated whether different coping responses are susceptible to renewal. During conditioning, male, Long-Evans rats were placed into a specific context (Context A) where an electrified shock-probe delivered a 3 mA shock upon contact. During extinction, the shock-probe was unarmed in either the same (Context A) or a different context (Context B). Renewal of conditioned responses was assessed in the conditioning context (ABA) or in a novel context (ABC or AAB). Renewal of passive coping responses, indicated by an increased latency and a decreased duration of shock-probe contacts, was observed in all groups. However, renewal of passive coping, measured by increased time spent on the side of the chamber opposite the shock-probe, was only found in the ABA group. Renewal of active coping responses linked to defensive burying was not observed in any group. The present findings highlight the presence of multiple psychological processes underlying even basic forms of aversive conditioning and demonstrate the importance of assessing a broader set of behaviours to tease apart these different underlying mechanisms. The current findings suggest that passive coping responses may be more reliable indicators for assessing renewal than active coping behaviours associated with defensive burying.

Impacts of Deoxygenation and Hypoxia on Shark Embryos Anti-Predator Behavior and Oxidative Stress.

Climate change is leading to the loss of oxygen content in the oceans and endangering the survival of many marine species. Due to sea surface temperature warming and changing circulation, the ocean has become more stratified and is consequently losing its oxygen content. Oviparous elasmobranchs are particularly vulnerable as they lay their eggs in coastal and shallow areas, where they experience significant oscillations in oxygen levels. Here, we investigated the effects of deoxygenation (93% air saturation) and hypoxia (26% air saturation) during a short-term period (six days) on the anti-predator avoidance behavior and physiology (oxidative stress) of small-spotted catshark (Scyliorhinus canicula) embryos. Their survival rate decreased to 88% and 56% under deoxygenation and hypoxia, respectively. The tail beat rates were significantly enhanced in the embryos under hypoxia compared to those exposed to deoxygenation and control conditions, and the freeze response duration showed a significant opposite trend. Yet, at the physiological level, through the analyses of key biomarkers (SOD, CAT, GPx, and GST activities as well as HSP70, Ubiquitin, and MDA levels), we found no evidence of increased oxidative stress and cell damage under hypoxia. Thus, the present findings show that the projected end-of-the-century deoxygenation levels elicit neglectable biological effects on shark embryos. On the other hand, hypoxia causes a high embryo mortality rate. Additionally, hypoxia makes embryos more vulnerable to predators, because the increased tail beat frequency will enhance the release of chemical and physical cues that can be detected by predators. The shortening of the shark freeze response under hypoxia also makes the embryos more prone to predation.

Anti-predator strategies of adult male Central Himalayan Langurs (Semnopithecus schistaceus) in response to domestic dogs in a human-dominated landscape.

The evolution of predator-prey relationships is an important topic in primatology. Many aspects of primate society have been explained as a response to predation pressure. While predation has been discussed in broad theoretical terms, few systematically collected data exist on the subject. Furthermore, little information exists regarding the inter-male variation in responses to predators. To address this data gap, predatory dog-primate interactions were studied in a 78-member group of habituated, individually recognized Central Himalayan Langurs (CHL) (Semnopithecus schistaceus) living in a high-altitude subsistence agricultural landscape of northern India. We recorded 312 langur-dog interactions over 2 years. These predation events resulted in 15 serious attacks on adult females, infants, juveniles and sub-adults, in eight of which the prey was killed and consumed on the spot. In response to dog predation, adult males performed three types of anti-predator response behaviors: direct fighting with a predator, emitting alarm calls, fleeing and/or freezing. Differences were noted in each male's response to village dogs. The results showed that the likelihood of CHL adult males engaging in more costly counterattacks or attention getting alarm calls were better predicted by the level of investment in the group (genetic relatedness, duration of residency, social relationships), but not rank and mating rate. Long-duration resident adult males performed high and/or intermediate cost behaviors to protect vulnerable members of the group; their potential offspring, maternal siblings or cousins, and adult female social partners. Short-term residents or recent immigrant males exhibited two less energetically costly, more self-preserving behaviors, depending on their rank: (1) high-ranking short-tenure duration males, with high mating frequencies, performed flee and freeze responses; (2) low-ranking, low-mating-frequency males performed more alarm calls. Counterattacks and alarm calls were performed by adult males with relatively more experience with village dogs and were directed towards dogs with predatory histories significantly more often than dogs with non-predatory histories. Natural selection and kin selection have both contributed to the evolution of CHL anti-predator tactics.

Perlite and Rice Husk Ash Re-Use As Fine Aggregates in Lightweight Aggregate Structural Concrete—Durability Assessment

In this paper, perlite mining and rice production by-products, namely run-of-mine perlite and rice husk ash, are used as fine aggregates in combination with pumice and calcareous aggregates to produce lightweight concrete. Their use is evaluated mainly in terms of the durability of the concrete, by comparing four optimized lightweight concrete mixtures of similar density and strength with a reference one of normal weight. The sorptivity due to capillary sorption, open porosity, chloride migration, penetration resistance, and freeze and thaw response were studied to evaluate the durability of the lightweight concrete. According to the experimental results, the examined mixtures developed an adequate strength in order to be classified into strength classes greater than LC25/28 and, therefore, be used in structural applications. The durability of the mixtures was also sufficient, especially as far as the chlorides’ penetration resistance is concerned, which was found to be up to 39% lower compared to the reference mixture. The sorptivity and open porosity of the LWC mixtures increased due to the porous nature of the lightweight aggregates, and the mixtures were also found to be susceptible to freeze and thaw cycles. Exceptionally, the lightweight concrete mixtures comprising pumice and perlite exhibited a lower sorptivity and resistance to chloride penetration than the standard concrete and a promising tolerance to freezing and thawing. Thus, the optimized combination of pumice and perlite is a sustainable recommendation for structural lightweight concrete production and use, promoting the wider exploitation of natural aggregates with an acceptable compromise on strength and durability.

Transcriptomics Reveals the Effect of Short-Term Freezing on the Signal Transduction and Metabolism of Grapevine.

Low temperature is an important factor limiting plant growth. Most cultivars of Vitis vinifera L. are sensitive to low temperatures and are at risk of freezing injury or even plant death during winter. In this study, we analyzed the transcriptome of branches of dormant cv. Cabernet Sauvignon exposed to several low-temperature conditions to identify differentially expressed genes and determine their function based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG)enrichment analyses. Our results indicated that exposure to subzero low temperatures resulted in damage to plant cell membranes and extravasation of intracellular electrolytes, and that this damage increased with decreasing temperature or increasing duration. The number of differential genes increased as the duration of stress increased, but most of the common differentially expressed genes reached their highest expression at 6 h of stress, indicating that 6 h may be a turning point for vines to tolerate extreme low temperatures. Several pathways play key roles in the response of Cabernet Sauvignon to low-temperature injury, namely: (1) the role of calcium/calmodulin-mediated signaling; (2) carbohydrate metabolism, including the hydrolysis of cell wall pectin and cellulose, decomposition of sucrose, synthesis of raffinose, and inhibition of glycolytic processes; (3) the synthesis of unsaturated fatty acids and metabolism of linolenic acid; and (4) the synthesis of secondary metabolites, especially flavonoids. In addition, pathogenesis-related protein may also play a role in plant cold resistance, but the mechanism is not yet clear. This study reveals possible pathways for the freezing response and leads to new insights into the molecular basis of the tolerance to low temperature in grapevine.

Behavioral and transcriptomic effects of the cancer treatment tamoxifen in mice.

Tamoxifen is a common treatment for estrogen receptor-positive breast cancer. While tamoxifen treatment is generally accepted as safe, there are concerns about adverse effects on cognition. We used a mouse model of chronic tamoxifen exposure to examine the effects of tamoxifen on the brain. Female C57/BL6 mice were exposed to tamoxifen or vehicle control for six weeks; brains of 15 mice were analyzed for tamoxifen levels and transcriptomic changes, and an additional 32 mice were analyzed through a battery of behavioral tests. Tamoxifen and its metabolite 4-OH-tamoxifen were found at higher levels in the brain than in the plasma, demonstrating the facile entry of tamoxifen into the CNS. Behaviorally, tamoxifen-exposed mice showed no impairment in assays related to general health, exploration, motor function, sensorimotor gating, and spatial learning. Tamoxifen-treated mice showed a significantly increased freezing response in a fear conditioning paradigm, but no effects on anxiety measures in the absence of stressors. RNA sequencing analysis of whole hippocampi showed tamoxifen-induced reductions in gene pathways related to microtubule function, synapse regulation, and neurogenesis. These findings of the effects of tamoxifen exposure on fear conditioning and on gene expression related to neuronal connectivity suggest that there may be CNS side effects of this common breast cancer treatment.

Compensatory Contextual Fear Memory Pathways Develop in the Infralimbic Cortex within 3 Days after the First Test in the Absence of the Dorsal Hippocampus.

The dorsal hippocampus (DH) is primarily involved in the formation of contextual fear-conditioned (CxFC) memory. However, CxFC memory can be formed even in the absence of the DH. In addition to the DH, the infralimbic cortex (IL), a sub-region of the medial prefrontal cortex (mPFC), also plays an important role in the consolidation of CxFC memory. However, role of IL in the development of compensatory CxFC memory is not known. Here, we have examined (a) the development of the compensatory circuitry of CxFC memory within 3 days after the first test in the absence of the DH and (b) the role of IL in the induction of compensatory CxFC memory in the absence of the DH. The DH-lesioned rats re-trained for CxFC 1 day after the first testing exhibited significantly less freezing compared to the control group. However, the DH-lesioned rats, re-trained for CxFC 3 days after the first testing, showed a robust freezing response. It suggests that the fully functional compensatory circuitry of contextual fear memory develops after multiple training separated by 3 days. Furthermore, we observed that reversible inactivation of the IL of the DH-lesioned rats during the first training waned the formation of compensatory CxFC. It suggests that (a) the IL receives contextual fear memory information during the first trial in the absence of the DH and (b) perturbation in fear memory information encoding in the IL during the first trial impairs the development of the compensatory network in the absence of the DH.

Abstract TP215: Prenatal Alcohol Exposure Impairs Long-term Behavioral Recovery Following Ischemic Stroke In Sprague Dawley Rats

Background and Purpose: Early life adversity is an important risk factor for cardiometabolic diseases in adulthood. There is very little information on prenatal insults and adult stroke outcomes. The present study tested the hypothesis that prenatal alcohol exposure (PAE) impedes long-term recovery from ischemic stroke in rats. Methods: Pregnant Sprague Dawley rats (in-house bred) were exposed to 10% alcohol or air, for 1 hour from gestational day 8 through 18 using the rodent alcohol inhalation system. Adult offspring (5-6 mo old) were subjected to endothelin-1 induced MCAo and outcomes were evaluated after 180 days. Long-term cognitive outcomes were determined using the Barnes maze and fear conditioning/extinction test. Results: In the Barnes maze test, control males showed shorter latency to reach the goal box over 3-day learning period (p=0.002), as assessed by the slope of the curve (-27.66, significantly different from 0; p=0.005). In contrast, the latency over the 3-day learning period did not improve in PAE males (-9.674, not significantly different from 0; p=0.22). In females, the latency to escape box during the learning trials was not different between the control and PAE. The fear conditioning data indicate that percent freezing over trials was similar in control and PAE-males. However, the PAE females exhibited a diminished freezing response as compared to control females (p=0.01). In the context of fear extinction, control males showed a reduction in freezing response to repeated tones over the trial blocks (p=0.004) compared to PAE-males. The control and PAE females showed similar freezing responses with repeated tones. Conclusions: The present findings support the hypothesis that PAE modifies long-term cognitive changes after stroke, and further show that different cognitive domains are affected by PAE in males and females.

THE FUTURE ROLE OF THE INTERNAL AUDIT FUNCTION: ASSURE. BUILD. CONSULT

Chambers (2022) recently raised “a red flag” by pointing out that internal auditors have been unduly placing Environmental, Social, and Governance (ESG) risks on the back burner. Internal auditors currently do not play a significant role as assurance providers and are absent from potential advisory services about ESG – on both sides of the Atlantic. We diagnose an “ESG helplessness syndrome.” Like in the world of animals, the internal audit function is in a state of freeze response when it comes to ESG topics. The ESG challenge is so big, and the threats for the role of the Internal Audit Function (IAF) are so real, that the profession reacts like animals in the face of a threat: they freeze. We discuss and challenge the professional demand for “objectivity” and “independence” in the ESG context as they might represent obstacles to the IAF playing a significant role in the ESG agenda. We suggest practitioners consider widening the repertoire of internal auditing. We suggest an ABC-Model © of Internal Auditing, adding “Building” as a new third pillar of internal audit value creation which complements the traditional assurance and consulting services. We encourage internal auditors to become “builders” when tackling the ESG challenge in their respective organizations. Metaphorically speaking, we borrow from Yvon Chouinard, the founder of Patagonia which is often used as an ESG role model company when we suggest “Let Internal Auditors Go Surfing” as our call to action.

Anatomical and functional study of the cuneiform nucleus: A critical site to organize innate defensive behaviors.

The cuneiform nucleus (CUN) is a midbrain structure located lateral to the caudal part of the periaqueductal gray. In the present investigation, we first performed a systematic analysis of the afferent and efferent projections of the CUN using FluoroGold and Phaseolus vulgaris leucoagglutinin as retrograde and anterograde neuronal tracers, respectively. Next, we examined the behavioral responses to optogenetic activation of the CUN and evaluated the impact of pharmacological inactivation of the CUN in both innate and contextual fear responses to a predatory threat (i.e., a live cat). The present hodologic evidence indicates that the CUN might be viewed as a caudal component of the periaqueductal gray. The CUN has strong bidirectional links with the dorsolateral periaqueductal gray (PAGdl). Our hodological findings revealed that the CUN and PAGdl share a similar source of inputs involved in integrating information related to life-threatening events and that the CUN provides particularly strong projections to brain sites influencing antipredatory defensive behaviors. Our functional studies revealed that the CUN mediates innate freezing and flight antipredatory responses but does not seem to influence the acquisition and expression of learned fear responses.

Effects of stress and mindfulness on epigenetics.

Epigenetics are heritable changes in the rate of gene expression without any modification of the DNA sequence and occur in response to environmental changes. Tangible changes to the external surroundings may be practical causes for epigenetic modifications, playing a potential evolutionary role. While fight, flight, or freeze responses once served a concrete role in survival, modern humans may not face similar existential threats that warrant psychological stress. Yet, chronic mental stress is predominant in modern life. This chapter elucidates the deleterious epigenetic changes that occur due to chronic stress. In an exploration of mindfulness-based interventions (MBIs) as a potential antidote to such stress-induced epigenetic modifications, several pathways of action are uncovered. The epigenetic changes that occur because of mindfulness practice are demonstrated across the hypothalamic-pituitary-adrenal axis, serotonergic transmission, genomic health and aging, and neurological biomarkers.

Frozen mountain pine needles: The endodermis discriminates between the ice-containing central tissue and the ice-free fully functional mesophyll.

Conifer (Pinaceae) needles are the most frost-hardy leaves. During needle freezing, the exceptional leaf anatomy, where an endodermis separates the mesophyll from the vascular tissue, could have consequences for ice management and photosynthesis. The eco-physiological importance of needle freezing behaviour was evaluated based on the measured natural freezing strain at the alpine treeline. Ice localization and cellular responses to ice were investigated in mountain pine needles by cryo-microscopic techniques. Their consequences for photosynthetic activity were assessed by gas exchange measurements. The freezing response was related to the microchemistry of cell walls investigated by Raman microscopy. In frozen needles, ice was confined to the central vascular cylinder bordered by the endodermis. The endodermal cell walls were lignified. In the ice-free mesophyll, cells showed no freeze-dehydration and were found photosynthetically active. Mesophyll cells had lignified tangential cell walls, which adds rigidity. Ice barriers in mountain pine needles seem to be realised by a specific lignification patterning of cell walls. This, additionally, impedes freeze-dehydration of mesophyll cells and enables gas exchange of frozen needles. At the treeline, where freezing is a dominant environmental factor, the elaborate needle freezing pattern appears of ecological importance. This article is protected by copyright. All rights reserved.

Depersonalisation – a key to understand the complexity of dissociative phenomena

The concepts of depersonalisation and derealisation refer to alteration in how an affected individual feels about oneself and their environment. Both these experiences may range from mild and episodic to severe and chronic. When clinically severe, these symptoms can be the basis for the diagnosis of depersonalisation-derealisation disorder; however, they are viewed as transdiagnostic, as they accompany various mental disorders. Literature emphasises the heterogeneous nature of the discussed construct, which includes several dimensions such as derealisation, somatopsychic depersonalisation, autopsychic depersonalisation and affective depersonalisation. This phenomenological complexity is reflected in the complex neurobiological mechanisms responsible for depersonalisation experiences, of which the theory of cortico-limbic inhibition is best described. Symptoms of depersonalisation and derealisation can be triggered by a variety of factors, such as physiological states (fatigue, stress, etc.), central nervous system abnormalities or psychoactive substances. In many cases, however, traumatic experiences are crucial, with most empirical evidence pointing to the role of emotional abuse and neglect. Trauma can alter responses of the autonomic nervous system, which mediates the symptoms of depersonalisation and derealisation (e.g. freeze response). The attachment style may be another mediator. Although depersonalisation and derealisation were conventionally assigned to the category of detachment, it has been suggested that they also bear features of compartmentalisation under certain circumstances. The analysis of the impact of various developmental processes on the formation of the complex phenomenon of depersonalisation sheds new light on the understanding of dissociative disorders and the importance of dissociation in psychopathology.

Social Behavioral Deficits in Krushinsky-Molodkina Rats, an Animal Model of Audiogenic Epilepsy.

In clinical practice, epilepsy is often comorbid with the autism spectrum disorders (ASDs). This warrants a search of animal models to uncover putative overlapping neuronal mechanisms. The Krushinsky-Molodkina (KM) rat strain is one of the oldest inbred animal models for human convulsive epilepsies. We analyzed the behavioral response of adult seizure-naive KM males in three-chambered tests for social preference. We found that a presence of social stimuli (encaged unfamiliar Wistar rats of the same age and sex) evoked a reduced or reversed exploratory response in freely moving KM individuals. The epilepsy-prone rats demonstrated remarkably shortened bouts of social contacts and displayed less locomotion around the stranger rat-containing boxes, together with a pronounced freezing response. The decrease in social preference was not due to a general decrease in activity, since relative measures of activity, the index of sociability, were decreased, too. The susceptibility to audiogenic seizures was verified in the KM cohort but not seen in the control Wistar group. We propose the KM rat strain as a new animal model for comorbid ASD and epilepsy.

Involvement of D2-like dopaminergic receptors in contextual fear conditioning in female rats: influence of estrous cycle.

Introduction: Dopamine has been increasingly recognized as a key neurotransmitter regulating fear/anxiety states. Nevertheless, the influence of sex and estrous cycle differences on the role of dopamine in fear responses needs further investigation. We aimed to evaluate the effects of sulpiride (a dopaminergic D2-like receptor antagonist) on contextual fear conditioning in females while exploring the influence of the estrous cycle. Methods: First, using a contextual fear conditioning paradigm, we assessed potential differences in acquisition, expression, and extinction of the conditioned freezing response in male and female (split in proestrus/estrus and metestrus/diestrus) Wistar rats. In a second cohort, we evaluated the effects of sulpiride (20 and 40 mg/kg) on contextual conditioned fear in females during proestrus/estrus and metestrus/diestrus. Potential nonspecific effects were assessed in motor activity assays (catalepsy and open-field tests). Results: No sex differences nor estrous cycle effects on freezing behavior were observed during the fear conditioning phases. Sulpiride reduced freezing expression in female rats. Moreover, females during the proestrus/estrus phases of the estrous cycle were more sensitive to the effects of sulpiride than females in metestrus/diestrus. Sulpiride did not cause motor impairments. Discussion: Although no sex or estrous cycle differences were observed in basal conditioned fear expression and extinction, the estrous cycle seems to influence the effects of D2-like antagonists on contextual fear conditioning.

Silence and related symptoms in children and adolescents: a network approach to selective mutism

BackgroundSilence in certain situations represents the core symptom of selective mutism (SM). However, it is unclear what additional symptoms are part of this disorder. Although knowledge of symptoms is essential for diagnostics and intervention, to date, only scarce research exists on circumscribed symptoms of SM. Given the large overlap between SM and social anxiety disorder (SAD), it remains also unclear which symptoms can differentiate both disorders.MethodsA network analysis of potential symptoms of SM was performed based on a mixed sample of N = 899 children and adolescents with and without indication of SM (n = 629 with silence in certain situations). In a preliminary analysis, we demonstrated that children with and without silence in certain situations do not differ with respect to their network structure, justifying an analysis on the entire mixed sample. Possible communities (symptom clusters) within the network and thus potential latent variables were examined, and symptoms were analyzed in terms of their centrality (the extent to which they are associated with other symptoms in the network). To investigate the differentiability of symptoms of the SM network from symptoms of SAD, we computed a network that additionally contains symptoms of SAD.ResultsIn the resulting network on symptoms of SM, silence was, as expected, the symptom with the highest centrality. We identified two communities (symptom cluster): (1) symptoms associated with the fear response of freezing, (2) symptoms associated with speech production and avoidance. SM network symptoms and SAD symptoms largely formed two separate symptom clusters, with only selectivity of speaking behavior (more talkative at home and taciturn or mute outside the home) falling into a common cluster with SAD symptoms.ConclusionsSilence appears to have been confirmed by analysis as a core symptom of SM. Additional anxiety-related symptoms, such as avoidance behavior or motor inhibition associated with freezing, seem to co-occur with silence. The two communities of SM potentially indicate different mechanisms of silence. The symptoms of SM appear to be distinguishable from those of SAD, although there seems to be overlap in terms of difficulty speaking in situations outside the home.

Identification of key gene networks related to the freezing resistance of apricot kernel pistils by integrating hormone phenotypes and transcriptome profiles

BackgroundApricot kernel, a woody oil tree species, is known for the high oil content of its almond that can be used as an ideal feedstock for biodiesel production. However, apricot kernel is vulnerable to spring frost, resulting in reduced or even no yield. There are no effective countermeasures in production, and the molecular mechanisms underlying freezing resistance are not well understood.ResultsWe used transcriptome and hormone profiles to investigate differentially responsive hormones and their associated co-expression patterns of gene networks in the pistils of two apricot kernel cultivars with different cold resistances under freezing stress. The levels of auxin (IAA and ICA), cytokinin (IP and tZ), salicylic acid (SA) and jasmonic acid (JA and ILE-JA) were regulated differently, especially IAA between two cultivars, and external application of an IAA inhibitor and SA increased the spring frost resistance of the pistils of apricot kernels. We identified one gene network containing 65 hub genes highly correlated with IAA. Among these genes, three genes in auxin signaling pathway and three genes in brassinosteroid biosynthesis were identified. Moreover, some hub genes in this network showed a strong correlation such as protein kinases (PKs)-hormone related genes (HRGs), HRGs-HRGs and PKs-Ca2+ related genes.ConclusionsCa2+, brassinosteroid and some regulators (such as PKs) may be involved in an auxin-mediated freezing response of apricot kernels. These findings add to our knowledge of the freezing response of apricot kernels and may provide new ideas for frost prevention measures and high cold–resistant apricot breeding.

TRPV1 modulation of contextual fear memory depends on stimulus intensity and endocannabinoid signalling in the dorsal hippocampus

The transient receptor potential vanilloid type-1 (TRPV1) channels have been implicated in the modulation of aversive responses. The endocannabinoid anandamide acts as an endogenous TRPV1 agonist, exerting opposite functions at TRPV1 and type-1 cannabinoid receptors (CB1R). Here we tested the hypothesis that hippocampal TRPV1 modulates contextual fear memory retrieval and investigated the influence of the aversive stimulus intensity as well as the role of endocannabinoid signaling. Male C57BL/6J mice were tested for contextual fear memory after low-, moderate-, or high-intensity shock protocols. The selective TRPV1 blockers SB366791 (1-10 nmol) and 6-I-NC (2 nmol) were infused via intra-dorsal hippocampus before the retrieval test session. The local levels of endocannabinoids and Arc and Zif268 mRNAs, involved in synaptic plasticity and memory, were quantified. First, both TRPV1 blockers reduced memory retrieval in animals exposed to moderate or high (but not low) intensity training protocols. In the second series of results, the magnitude of the freezing responses positively correlated with the hippocampal anandamide levels; TRPV1 and CB1R were found co-localized in this brain region; and the CB1R antagonist, AM251, prevented the effects of SB366791. Thus, endocannabinoid signaling possibly mediates the effects of TRPV1 blockers. Finally, inhibition of memory retrieval by TRPV1 blockers increased Arc and Zif268 mRNAs and impaired fear memory reinstatement. In conclusion, the modulation of fear memories by dorsal hippocampal TRPV1 channels may depend on the aversive stimulus intensity and occur via anandamide/CB1 signaling. Moreover, TRPV1 blockers promote Arc and Zif268 transcription, with subsequent attenuation of aversive memory reinstatement.

Re-exposures in the Dark Cycle Promote Attenuation of Fear Memory: Role of the Circadian Cycle and Glucocorticoids

It has been shown that a previously consolidated memory can incorporate either new external information or a novel internal emotional state following a labile state induced by retrieval. This updating process allows editing unwanted fear memory, leading to the reduction of the fear response. Memory can be modulated by the circadian cycle. Considering that rodents are more active during the night, expressing less fearful behavior, we investigated whether fear memory can be updated when reactivated during the dark cycle. We found that rats expressed lower freezing levels during a single retrieval session in the dark cycle, but not in the test. However, three retrieval sessions in the dark cycle were able to update fear memory, reducing freezing response in the test performed in the light cycle. This effect was blocked when the glucocorticoid synthesis inhibitor metyrapone was administered before retrieval. This approach opens new avenues to explore interventions that consider the circadian cycle in the treatment of fear memories based on non-pharmacological interventions.

How resistant are levodopa-resistant axial symptoms? Response of freezing, posture, and voice to increasing levodopa intestinal infusion rates in Parkinson disease.

Treatment of freezing of gait (FoG) and other Parkinson disease (PD) axial symptoms is challenging. Systematic assessments of axial symptoms at progressively increasing levodopa doses are lacking. We sought to analyze the resistance to high levodopa doses of FoG, posture, speech, and altered gait features presenting in daily-ON therapeutic condition. We performed a pre-/postinterventional study including patients treated with levodopa/carbidopa intestinal gel infusion (LCIG) with disabling FoG in daily-ON condition. Patients were evaluated at their usual LCIG infusion rate (T1), and 1 h after 1.5× (T2) and 2× (T3) increase of the LCIG infusion rate by quantitative outcome measures. The number of FoG episodes (primary outcome), posture, speech, and gait features were objectively quantified during a standardized test by a blinded rater. Changes in motor symptoms, dyskinesia, and plasma levodopa concentrations were also analyzed. We evaluated 16 patients with a mean age of 69 ± 9.4 years and treated with LCIG for a mean of 2.2± 2.1 years. FoG improved in 83.3% of patients by increasing the levodopa doses. The number of FoG episodes significantly decreased (mean=2.3 at T1, 1.7 at T2, 1.2 at T3; p= 0.013). Posture and speech features did not show significant changes, whereas stride length (p= 0.049), turn duration (p= 0.001), and turn velocity (p= 0.024) significantly improved on doubling the levodopa infusion rate. In a short-term evaluation, the increase of LCIG dose can improve "dopa-resistant" FoG and gait issues in most advanced PD patients with overall good control of motor symptoms in the absence of clinically significant dyskinesia.