Correction: Data-driven analyses of motor impairments in animal models of neurological disorders.

[This corrects the article DOI: 10.1371/journal.pbio.3000516.].

Regional Differences in BDNF Expression and Behavior as a Function of Sex and Enrichment Type: Oxytocin Matters.

The early environment is critical to brain development, but the relative contribution of physical versus social stimulation is unclear. Here, we investigated in male and female rats the response to early physical and social environmental enrichment in relation to oxytocin (OT) and brain-derived neurotrophic factor (BDNF) expression. The findings show that males and females respond differently to prolonged sensorimotor stimulation from postnatal days 21-110 in terms of functional, structural, and molecular changes in the hippocampus versus medial prefrontal cortex (mPFC). Physical enrichment promoted motor and cognitive functions and hippocampal BDNF mRNA and protein expression in both sexes. Combined physical and social enrichment, however, promoted functional and structural gain in females. These changes were accompanied by elevated plasma oxytocin (OT) levels and BDNF mRNA expression in the mPFC, while the hippocampus was not affected. Administration of an OT antagonist in females blocked the beneficial effects of enrichment and led to reduced cortical BDNF signaling. These findings suggest that an OT-based mechanism selectively stimulates a region-specific BDNF response which is dependent on the type of experience.

Environmental Enrichment Promotes Transgenerational Programming of Uterine Inflammatory and Stress Markers Comparable to Gestational Chronic Variable Stress.

Prenatal maternal stress is linked to adverse pregnancy and infant outcomes, including shortened gestation lengths, low birth weights, cardio-metabolic dysfunction, and cognitive and behavioural problems. Stress disrupts the homeostatic milieu of pregnancy by altering inflammatory and neuroendocrine mediators. These stress-induced phenotypic changes can be passed on to the offspring epigenetically. We investigated the effects of gestational chronic variable stress (CVS) in rats using restraint and social isolation stress in the parental F0 generation and its transgenerational transmission across three generations of female offspring (F1-F3). A subset of F1 rats was housed in an enriched environment (EE) to mitigate the adverse effects of CVS. We found that CVS is transmitted across generations and induces inflammatory changes in the uterus. CVS did not alter any gestational lengths or birth weights. However, inflammatory and endocrine markers changed in the uterine tissues of stressed mothers and their offspring, suggesting that stress is transgenerationally transmitted. The F2 offspring reared in EE had increased birth weights, but their uterine gene expression patterns remained comparable to those of stressed animals. Thus, ancestral CVS induced changes transgenerationally in fetal programming of uterine stress markers over three generations of offspring, and EE housing did not mitigate these effects.

Data-driven analyses of motor impairments in animal models of neurological disorders.

Behavior provides important insights into neuronal processes. For example, analysis of reaching movements can give a reliable indication of the degree of impairment in neurological disorders such as stroke, Parkinson disease, or Huntington disease. The analysis of such movement abnormalities is notoriously difficult and requires a trained evaluator. Here, we show that a deep neural network is able to score behavioral impairments with expert accuracy in rodent models of stroke. The same network was also trained to successfully score movements in a variety of other behavioral tasks. The neural network also uncovered novel movement alterations related to stroke, which had higher predictive power of stroke volume than the movement components defined by human experts. Moreover, when the regression network was trained only on categorical information (control = 0; stroke = 1), it generated predictions with intermediate values between 0 and 1 that matched the human expert scores of stroke severity. The network thus offers a new data-driven approach to automatically derive ratings of motor impairments. Altogether, this network can provide a reliable neurological assessment and can assist the design of behavioral indices to diagnose and monitor neurological disorders.

Looking beyond the standard version of the Morris water task in the assessment of mouse models of cognitive deficits.

Most studies investigating hippocampal-dependent learning and memory in mouse models of disease use the standard version of the Morris water task (MWT), in which a place is learned over several days. While useful in determining if there are learning and memory deficits, often it is not clear if memory acquisition, consolidation, or retrieval is affected. For rats, we developed a variant of the task in which we added a single-massed training session to a new location after the standard distributed version of the MWT. Using this version of the task, competition between these two spatial representations can then be assessed in a probe trial. We have found in rat models of Alzheimer's disease that this paradigm can detect subtle impairments that are often missed in the standard version of the MWT. To the best of our knowledge, MWT paradigm with a single-massed training session have never been used for mice. We sought to validate this paradigm for the use of assessing mouse models of disease. In the first two experiments, control mice did not have a preference for the new platform location, but instead with extensive training in the massed session displayed a preference for both the old and new locations. In the third experiment, a novel mouse model of Alzheimer's disease was impaired in the standard version of the MWT, but not in the massed training phase of this paradigm. Importantly, these data demonstrate that our paradigm is more informative in characterizing spatial learning and memory in mouse models of disease.

Protective effect of raisin (currant) against spatial memory impairment and oxidative stress in Alzheimer disease model.

Objectives: Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive pathological changes of the brain. A number of studies demonstrated compelling evidence of the importance of oxidative processes in AD pathogenesis. Raisin contains polyphenol, phenolic acid, and tannin compounds, which have antioxidant and anti-inflammatory properties. The present study was aimed to evaluate the protective effect of raisin on neurobehavioral and histological changes in rats with Alzheimer. Methods: Animal model of AD was induced by intraperitoneal injection of aluminium chloride for 60 days (100 mg/kg body weight). During these 60 days both Alzheimer's and control rats were given 6 g of raisin per rat. At the end of the treatment, blood was collected for biochemical assessment. We used a Morris water task and passive avoidance test to assess spatial memory. Results: Our results showed that aluminium exposure significantly decreased the memory in the MWT and passive avoidance test, but in the raisin + AlCl3 group, it significantly increased spatial memory in both tests. Also, Aluminium exposure significantly increased malondialdehyde (MDA) and decreased ferric reducing ability of plasma (ferric reducing/antioxidant power (FRAP)), while treatment with raisin significantly decreased MDA and increased FRAP in plasma of blood. Discussion: Our findings showed that raisin has a neuroprotective effect and improves the spatial memory in AD animal models.

Organization of the reach and grasp in head-fixed vs freely-moving mice provides support for multiple motor channel theory of neocortical organization.

Multiple motor channel (MMC) theory of neocortical organization proposes that complex movements, such as reaching for a food item to eat, are produced by the coordinated action of separate neural channels. For example, the human reach-to-grasp act is mediated by two visuo-parieto-motor cortex channels, one for the reach and one for the grasp. The present analysis asked whether there is a similar organization of reach-and-grasp movements in the mouse. The reach-to-eat movements of the same mice were examined from high-shutter speed, frame-by-frame video analysis in three tasks in which the mice obtained equivalent success scores: when freely-moving reaching for food pellets, when head-fixed reaching for food pellets, and when head-fixed reaching for pieces of pasta. To reach, the mice used egocentric cues to vary upper arm movements in a task-appropriate manner to place an open hand on the food or to locate the food using a "touch-release-grasp" strategy. Although mice could not hand-shape offline when reaching, they could hand-shape using online touch-related cues from the mouth to manipulate the food at the mouth. That the reach can be performed offline in relation to egocentric cues whereas hand shaping for the grasp requires online cues supports the idea that for the mouse, as for primates, the reach and grasp are separate acts. The results are further discussed in relation to the use of the head-fixed behavioral procedure to identify the independent neural substrates of the reach and the grasp using mesoscale stimulation/imaging methods.

Stress inhibits psychomotor performance differently in simple and complex open field environments.

Stress affects psychomotor profiles and exploratory behavior in response to environmental features. Here we investigated psychomotor and exploratory patterns induced by stress in a simple open-field arena and a complex, multi-featured environment. Groups of rats underwent seven days of restraint stress or no-stress conditions and were individually tested in three versions of the ziggurat task (ZT) that varied according to environmental complexity. The hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis due to stress procedure was evaluated by the pre- and post-stress levels of circulating corticosterone (CORT). Horizontal activity, exploration, and motivation were measured by the number of fields entered, the time spent in the central fields, path length and speed, and stop duration. In addition, vertical exploratory behavior was measured by the times rats climbed onto ziggurats. Stress-induced psychomotor changes were indicated by reduced path length and path speed and increased duration of stops only within the complex arena of the ZT. Rats in stress groups also showed a significant decline in the vertical movements as measured by the number of climbing onto ziggurats. No stress-induced changes were revealed by the simple open-field arena. The exploratory patterns of stressed animals suggest psychomotor inhibition and reduced novelty-seeking behaviors in an environment-dependent manner. Thus, multi-featured arenas that require complex behavioral strategies are ideally suited to reveal the inhibitory effects of stress on psychomotor capabilities in rodents.

Toward reframing brain-social dynamics: current assumptions and future challenges.

Evolutionary analyses suggest that the human social brain and sociality appeared together. The two fundamental tools that accelerated the concurrent emergence of the social brain and sociality include learning and plasticity. The prevailing core idea is that the primate brain and the cortex in particular became reorganised over the course of evolution to facilitate dynamic adaptation to ongoing changes in physical and social environments. Encouraged by computational or survival demands or even by instinctual drives for living in social groups, the brain eventually learned how to learn from social experience via its massive plastic capacity. A fundamental framework for modeling these orchestrated dynamic responses is that social plasticity relies upon neuroplasticity. In the present article, we first provide a glimpse into the concepts of plasticity, experience, with emphasis on social experience. We then acknowledge and integrate the current theoretical concepts to highlight five key intertwined assumptions within social neuroscience that underlie empirical approaches for explaining the brain-social dynamics. We suggest that this epistemological view provides key insights into the ontology of current conceptual frameworks driving future research to successfully deal with new challenges and possible caveats in favour of the formulation of novel assumptions. In the light of contemporary societal challenges, such as global pandemics, natural disasters, violent conflict, and other human tragedies, discovering the mechanisms of social brain plasticity will provide new approaches to support adaptive brain plasticity and social resilience.

Early life stress aggravates disease pathogenesis in mice with experimental autoimmune encephalomyelitis: Support for a two-hit hypothesis of multiple sclerosis etiology.

Vision problems are one of the earliest diagnosed symptoms of multiple sclerosis (MS). The onset and progression of vision loss and the underlying pathogenesis in MS may be influenced by cumulative psychophysiological stress. Here, we used a two-hit model of stress in female mice to determine if early life stress (ELS, the first hit) influences the response to an immunization that induces experimental autoimmune encephalomyelitis (EAE, the second hit) later in life. We hypothesized that ELS caused by animal transportation from a vendor during early postnatal development represents a co-factor which can exacerbate the clinical severity of EAE. Indeed, adult EAE mice with a history of ELS displayed more severe clinical signs and delayed recovery compared to non-stressed EAE mice. ELS also diminished visual acuity measured by optokinetic responses, as well as locomotion and exploratory behaviours in EAE mice. Notably, ELS accelerated vision loss and caused earlier onset of visual impairments in EAE. Exacerbated functional impairments in stressed EAE mice were highly correlated with circulating corticosterone levels. The findings show that the progression of induced EAE in adulthood can be significantly impacted by adverse early life experiences. These observations emphasize the importance of comprehensive behavioural testing, including non-motor functions, to enhance the translational value of preclinical animal models of MS. Moreover, shipment stress of laboratory animals should be considered a necessary variable in preclinical MS research. The consideration of cumulative lifetime stresses provides a new perspective of MS pathogenesis within a personalized medicine framework.

Thermoregulatory dynamics reveal sex-specific inflammatory responses to experimental autoimmune encephalomyelitis in mice: Implications for multiple sclerosis-induced fatigue in females.

The course of multiple sclerosis (MS) is characterized by striking sex differences in symptoms such as fatigue and impaired thermal regulation, which are associated with aggravated systemic pro-inflammatory processes. The purpose of this study was to replicate these symptoms in experimental autoimmune encephalomyelitis (EAE) in C57BL/6 mice in the quest to advance the preclinical study of non-motor symptoms of MS. Male and female C57BL/6 mice exposed to a mild form of EAE were evaluated for the progression of clinical, behavioural, thermal, and inflammatory processes. We show higher susceptibility in females to EAE than males based on greater clinical score and cumulative disease index (CDI), fatigue-like and anxiety-like behaviours. Accordingly, infrared (IR) thermography indicated higher cutaneous temperatures in females from post-induction days 12-23. Females also responded to EAE with greater splenic and adrenal gland weights than males as well as sex-specific changes in pro- and anti-inflammatory cytokines. These findings provide the first evidence of a sex-specific thermal response to immune-mediated demyelination, thus proposing a non-invasive assessment approach of the psychophysiological dynamics in EAE mice. The results are discussed in relation to the thermoregulatory correlates of fatigue and how endogenously elevated body temperature without direct heat exposure may be linked to psychomotor inhibition in patients with MS.

Bladder dysfunction in experimental autoimmune encephalomyelitis reflects clinical severity: A pilot study.

Multiple sclerosis (MS) is commonly associated with bladder dysfunction resulting in a progressive loss of voluntary control for urination over time. Here, we used the voided stain on paper (VSOP) method to investigate bladder function in the experimental autoimmune encephalomyelitis (EAE) mouse model of MS. Using the VSOP test, we show that bladder dysfunction reflects pro-inflammatory processes of EAE and severity of clinical EAE symptoms, as characterized by increased urine voided volume per micturition (UVVM) on post-induction day 7 and decreased UVVM on post-induction day 14. The UVVM was closely related to a clinical disease index of EAE symptoms and plasma granulocyte-macrophage colony-stimulating factor (GM-CSF) cytokine levels. UVVM was also sensitive to early life stress caused by animal transportation, which diminished UVVM at the peak of symptoms on post-induction day 14 in EAE mice. The results indicate that symptoms and progression of EAE can be reliably measured by VSOP as a non-motor function assessment.

Sex-specific stress and biobehavioral responses to human experimenters in rats.

Important factors influencing the outcome of animal experiments in preclinical research are often overlooked. In the current study, the reaction of female and male rats toward the biological sex of a human experimenter was investigated in terms of anxiety-like behaviors and physiological stress responses, as measured by infrared (IR) thermography, circulating corticosterone (CORT) and oxytocin levels. Female rats displayed consistently exacerbated anxiety-related behaviors along with elevated body surface temperature during repeated exposure to male experimenters. Experimental stress further intensified thermal responses to a male experimenter, especially in female rats. The behavioral responses to a male experimenter in females were associated with higher circulating CORT and lower oxytocin levels. Similar responses were induced by a T-shirt worn by a human male. The findings suggest that psychophysiological responses of female rats to a male experimenter are influenced by both visual and olfactory cues. The results emphasize the need to not only consider sex differences in experimental animals, but also standardize and report the experimenter's biological sex to avoid ambiguity in the generation and interpretation of results.

Chronic stress prior to hippocampal stroke enhances post-stroke spatial deficits in the ziggurat task.

Stress is one of the most important variables to determine recovery following stroke. We have previously reported that post-stroke exposure to either stress or corticosterone (CORT) alleviates hippocampal ischemic outcome. The present experiment expands previous findings by investigating the influence of exposure to stress prior to ischemic event. Rats received either daily restraint stress (1h/day; 16 consecutive days) or CORT (0.5mg/kg; 16 consecutive days) prior to focal ischemic stroke in the hippocampus induced by bilateral injection of endothelin-1 (ET-1). All experimental groups were then tested in the ziggurat task, a new task for spatial cognition. The stress+stroke group showed significant deficits in both hippocampal structure and function. No deleterious effect of pre-stroke exposure to CORT was found in the CORT+stroke group. Our results indicate that a history of chronic stress sensitizes hippocampal cells to the damaging consequences of focal ischemia. The opposing effects of CORT-related experiences in this study not only reflect the diversity of glucocorticoid actions in the stress response, but also provide evidence that elevated CORT in the absence of emotional disturbance is not sufficient to produce hippocampal deficit.

Aging, Social Distancing, and COVID-19 Risk: Who is more Vulnerable and Why?

Perceived social support represents an important predictor of healthy aging. The global COVID-19 pandemic has dramatically changed the face of social relationships and revealed elderly to be particularly vulnerable to the effects of social isolation. Social distancing may represent a double-edged sword for older adults, protecting them against COVID-19 infection while also sacrificing personal interaction and attention at a critical time. Here, we consider the moderating role of social relationships as a potential influence on stress resilience, allostatic load, and vulnerability to infection and adverse health outcomes in the elderly population. Understanding the mechanisms how social support enhances resilience to stress and promotes mental and physical health into old age will enable new preventive strategies. Targeted social interventions may provide effective relief from the impact of COVID-19-related isolation and loneliness. In this regard, a pandemic may also offer a window of opportunity for raising awareness and mobilizing resources for new strategies that help build resilience in our aging population and future generations.

Infrared Thermography Reveals Sex-Specific Responses to Stress in Mice.

Psychogenic hyperthermia is a stress-related condition reported mostly in women. Neuroendocrine responses to stress in females differ from those in males, and these differences cannot be explained solely based on hypothalamic-pituitary-adrenal (HPA) axis activity. Here, we used infrared (IR) thermographic imaging to record changes in cutaneous temperature following two types of stressful experiences in female and male mice. Mice were exposed to either single-session restraint stress or vertical exploration (rearing) deprivation and were monitored for exploratory activity and IR surface thermal changes. Females displayed higher rearing activity than males during the dark phase of the light cycle. Both sexes showed similar plasma corticosterone (CORT) responses after a challenge with restraint and rearing deprivation. However, only females responded to rearing deprivation with increased cutaneous temperature in the head and back, and a reduced thermal response in the tail. Circulating CORT levels were not correlated with the thermal variations. These findings, for the first time, provide evidence for sex-specific cutaneous thermal responses to short-term stress in mice following transient vertical-activity deprivation that may mimic clinical psychogenic hyperthermia.

Environmental determinants of behavioural responses to short-term stress in rats: Evidence for inhibitory effect of ambient landmarks.

Behavioural responses to stress occur in an environment-dependent manner. Complex environments require flexible behavioural coping strategies and chronic stress usually generates psychomotor inhibition. Here, we examine if short-term stress also exerts an inhibitory effect on novelty-seeking, exploratory behaviours. Rats underwent acute restraint stress or were left undisturbed, and their neuroendocrine and behavioural responses were assessed at short- and long-term time points. Animals were individually tested in the open field task (OFT) and the corridor field task (CFT) with and without a central object for free exploration and novelty seeking behaviour. Stress-related psychomotor alterations were measured by path speed, path length, number of stops and thigmotaxis in both tasks. Short-term stress activated the hypothalamic-pituitary-adrenal axis causing elevated plasma corticosterone levels. Stress also impacted psychomotor functions in terms of motivational changes (higher speed and longer path) only in the central-object variations of the OFT and CFT. Moreover, stress-induced emotional alterations were manifested by a higher number of stops and thigmotactic behaviour only in the central-object condition. These findings suggest that environmental landmarks determine the type and direction of exploratory behaviour under transient stress.

Rats with hippocampal lesion show impaired learning and memory in the ziggurat task: a new task to evaluate spatial behavior.

Spatial tasks are widely used to determine the function of limbic system structures in rats. The present study used a new task designed to evaluate spatial behavior, the ziggurat task (ZT), to examine the performance of rats with widespread hippocampal damage induced by N-methyl-d-aspartic acid (NMDA). The task consisted of an open field containing 16 identical ziggurats (pyramid shaped towers) arranged at equal distances. One of the ziggurats was baited with a food reward. The task required rats to navigate through the open field by using a combination of distal and/or proximal cues in order to locate the food reward. The ability to acquire and recall the location of the goal (baited) ziggurat was tested in consecutive training sessions of eight trials per day for 10 days. The location of the goal ziggurat was changed every second day, requiring the rats to learn a total of five different locations. Several parameters, including latency to find the target, distance traveled, the number of visits to non-baited ziggurats (errors), and the number of returns were used as indices of learning and memory. Control rats showed a significant decrease in distance traveled and reduced latency in locating the goal ziggurat across trials and days, suggesting that they learned and remembered the location of the goal ziggurat. Interestingly, the hippocampal-damaged group moved significantly faster, and traveled longer distances compared to the control group. Significant differences were observed between these groups with respect to the number of errors and returns on test days. Day 11 served as probe day, in which no food reward was given. The controls spent more time searching for the food in the previous training quadrant compared to the hippocampal group. The findings demonstrate that the ZT is a sensitive and efficient dry task for measuring hippocampus-dependent spatial performance in rats requiring little training and not associated with some of the disadvantages of water tasks.

Spatial Memory and Antioxidant Protective Effects of Raisin (Currant) in Aged Rats.

Diets rich in fruits and vegetables can prevent age-related diseases. This research was conducted to evaluate the effects of raisin consumption on the spatial memory and morphometric parameters of brain tissue in aging rats. Old rats (20 months of age) were divided into 2 groups: control and raisin, with 6 rats in each group. The raisin group received 6 g of raisins daily in addition to their food and water for 90 days. After treatments, all animals were evaluated by behavioral tests to assess spatial memory and learning alongside other tests including the ferric reducing antioxidant power (FRAP), malondialdehyde, and histological examinations. The results showed that there are significant differences in the Morris water task and passive avoidance learning of behavioral tests and biochemical tests (FRAP and thiobarbituric acid reactive substances) between the two groups. The histological study indicated that the cell count of the hippocampus, the diameter of the lateral ventricle, and area of the corpus callosum in the raisin group changed in comparison with the control group but they were not significant. The results demonstrated that raisins significantly raise antioxidant levels in blood and promotes cognitive and motor performance in aging rats.

Non-diagnostic symptoms in a mouse model of autism in relation to neuroanatomy: the BTBR strain reinvestigated.

Several mouse models of autism spectrum disorder (ASD), including the BTBR T + tf/J (BTBR) inbred strain, display a diverse array of behavioral deficits with particular face validity. Here we propose that phenotyping these preclinical models of ASD should avoid excessive reliance on appearance validity of the behavioral observations. BTBR mice were examined in three non-diagnostic symptoms modalities, beside an anatomical investigation for construct validity. The BTBR strain displayed poor sensorimotor integration as reflected by shorter stride length and greater latency on the balance beam task (BBT) when compared with C57BL/6 (B6) controls. Also, locomotor indices in the open-field task (OFT) revealed that BTBR mice traveled longer distances with a remarkably faster exploration than the B6 group in favor of hyperactivity and impulsiveness. Furthermore, analysis of spatial performance including search strategies in the Morris water task (MWT) indicated spatial impairment in the BTBR strain due to failure to employ spatial strategies during navigation. Quantitative cytoarchitectonics and volumetric examinations also indicated abnormal cortical and subcortical morphology in the BTBR mice. The results are discussed in relation to the neuroanatomical correlates of motor and cognitive impairments in the BTBR strain. We conclude that non-diagnostic autistic-like symptoms in the BTBR mouse strain can be impacted by autism risk factors in a similar way than the traditional diagnostic signs.