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The study of circadian rhythms has been critically dependent upon analysing mouse home cage activity, typically employing wheel running activity under different lighting conditions. Here we assess a novel method, the Digital Ventilated Cage (DVC®, Tecniplast SpA, Italy), for circadian phenotyping. Based upon capacitive sensors mounted under black individually ventilated cages with inbuilt LED lighting, each cage becomes an independent light-controlled chamber. Home cage activity in C57BL/6J mice was recorded under a range of lighting conditions, along with circadian clock-deficient cryptochrome-deficient mice (Cry1−/−, Cry2−/− double knockout). C57BL/6J mice exhibited a 24 h period under light/dark conditions, with a free-running period of 23.5 h under constant dark, and period lengthening under constant light. Animals displayed expected phase shifting responses to jet-lag and nocturnal light pulses. Sex differences in circadian parameters and phase shifting responses were also observed. Cryptochrome-deficient mice showed subtle changes in activity under light/dark conditions and were arrhythmic under constant dark, as expected. Our results show the suitability of the DVC system for circadian behavioural screens, accurately detecting circadian period, circadian disruption, phase shifts and mice with clock defects. We provide an evaluation of the strengths and limitations of this method, highlighting how the use of the DVC for studying circadian rhythms depends upon the research requirements of the end user. [ABSTRACT FROM AUTHOR]

Introduction: Lung cancer is the first cause of cancer death in the world, due to a delayed diagnosis and the absence of efficacy therapies. KRAS mutation occurs in 25% of all lung cancers and the concomitant mutations in LKB1 determine aggressive subtypes of these tumors. The improvement of therapeutical options for KRASG12C mutations has increased the possibility of treating these tumors, but resistance to these therapies has emerged. Preclinical animal models permit the study of tumors and the development of new therapies. The DVC system was used to measure circadian activity changes indicative of lung cancer progression in KRAS and KRAS-LKB1 transgenic mouse models. Material and methods: KRAS and KRAS-LKB1 conditional transgenic animal models were bred and genotyped. The tumors were inducted using adeno-CRE-recombinase system. The mice were housed in a Digital Ventilated Cage (DVC®) rack measuring the locomotor activity continuously for 24/7. The progression of the tumors was monitored with MRI. The DVC system evaluated a reduction in animal locomotion during the tumor progression. Results: KRAS and KRAS-LKB1 mutations were induced, and the tumor formation and progression were monitored over time. As expected, the onset of the tumors in the two different breeds occurred at different times. DVC system registered the locomotion activity of the mice during the light and dark phases, reporting a strong reduction, mainly, in the dark phase. In KRAS-LKB1 models, the locomotion reduction appeared more pronounced than in KRAS models. Discussions: Transgenic animal models represent a fundamental tool to study the biology of cancers and the development of new therapies. The tumors induced in these models harbor the same genotypical and phenotypical characteristics as their human counterparts. DVC methods permit a home cage monitoring system useful for tracking animal behavior continuously 24 hours a day, 7 days a week. DVC system could determine disease progression by monitoring a single animal activity in a cage and also using group-housed animals. For these reasons, the DVC system could play a crucial role in identifying diseases at early stages and in testing new therapeutic approaches with a higher likelihood of efficacy. [ABSTRACT FROM AUTHOR]

Introduction: Our understanding of laboratory animal behavior and the implications of husbandry activities on their wellbeing remains incomplete. This is especially relevant with an aging colony as their activity patterns may change as they mature. Home Cage Monitoring (HCM) provides valuable insights into mouse activity within the animal's own environment and can shed light on acclimatization periods and responses to husbandry activities such as cage changing. The aim of this study was to monitor and explore changes in the activity and rest disturbance (RDI) patterns of an aging colony of male and female C57/BL6 mice. Methods: The mice were housed in the Digitally Ventilated Cage® system, for up to 18 months of age. Data was then downloaded to investigate how the activity patterns and RDI of the mice changed over time. Habituation, aging and cage change assessments were conducted using linear mixed models, while cage separation and stereotypic behavior investigations were conducted by visual inspection of the data. Results: As expected during the study, mice were less active during the light phase compared to the dark phase. However, on arrival mice displayed heightened activity and RDI during the light phase and reduced activity and RDI during the dark phase, taking several days to adjust to baseline "acclimatized" patterns. With age, overall activity significantly decreased from 5 months until 14 months of age, after which it increased back toward baseline levels. We also observed activity spikes during our monitoring of this colony. Prolonged housing can lead to alarming stereotypic behaviors in animals. Cages of mice flagged for potential stereotypy displayed sustained activity spikes in the light and dark phases. Spikes in activity during the dark phase were much more pronounced than in the light phase. Cage changing led to an increase in the light phase activity and RDI compared to the previous day, with no observed difference in the dark phase post-cage change. This effect remained consistent as the animals aged. Discussion: This study explores changes in the activity patterns of an aging colony of male and female C57/BL6 mice housed in the Digitally Ventilated Cage® system. We identified distinct aging phases concerning activity and RDI differences and a potential new welfare application for the DVC®, specifically for early detection of stereotypy. In conclusion, the adoption of HCM systems should be considered for long-term animal housing from both a welfare and behavioral perspective. [ABSTRACT FROM AUTHOR]

Locomotor activity can serve as a readout to identify discomfort and pain. Therefore, monitoring locomotor activity following interventions that induce potential discomfort may serve as a reliable method for evaluating animal health, complementing conventional methods such as body weight measurement. In this study, we used the digital ventilated cage (DVC®) system for the assessment of circadian locomotor activity, in addition to body weight monitoring, following intracranial stereotaxic surgery in an Alzheimer's disease mouse model (C57BL/6J/APPswe/PSEN1dE9). Stereotaxic surgery did not affect the organization of circadian locomotor activity of both 7-8-week-old and 19-21-week-old mice. However, we observed that both young and old mice exhibited a significant decrease in activity during the dark phase. Also, our study shows that changes in locomotor activity exhibit higher sensitivity in detecting alterations indicative of animal health compared to measuring body weight. In contrast to 7-8-week-old mice, where we observed no genotypic differences in locomotor activity, 19-21-week-old APP/PS1 mice showed increased locomotor activity compared to wild-type mice. Furthermore, our analyses revealed that a subset of the 7-8-week-old mice showed increased locomotor activity during the initial peak of the dark phase. One mouse experienced sudden death early in life, possibly due to epileptic seizures. Altogether, our findings affirm continuous activity measurements as used in the DVC® as a highly valuable objective method for post-surgical welfare monitoring. Its discerning capacity not only facilitates circadian locomotor rhythm assessment but also enables the identification of individual aberrant activity patterns, possibly indicative of epileptic seizures. [ABSTRACT FROM AUTHOR]

IntroductionLung cancer is the first cause of cancer death in the world, due to a delayed diagnosis and the absence of efficacy therapies. KRAS mutation occurs in 25% of all lung cancers and the concomitant mutations in LKB1 determine aggressive subtypes of these tumors. The improvement of therapeutical options for KRASG12C mutations has increased the possibility of treating these tumors, but resistance to these therapies has emerged. Preclinical animal models permit the study of tumors and the development of new therapies. The DVC system was used to measure circadian activity changes indicative of lung cancer progression in KRAS and KRAS-LKB1 transgenic mouse models.Material and methodsKRAS and KRAS-LKB1 conditional transgenic animal models were bred and genotyped. The tumors were inducted using adeno-CRE-recombinase system. The mice were housed in a Digital Ventilated Cage (DVC®) rack measuring the locomotor activity continuously for 24/7. The progression of the tumors was monitored with MRI. The DVC system evaluated a reduction in animal locomotion during the tumor progression.ResultsKRAS and KRAS-LKB1 mutations were induced, and the tumor formation and progression were monitored over time. As expected, the onset of the tumors in the two different breeds occurred at different times. DVC system registered the locomotion activity of the mice during the light and dark phases, reporting a strong reduction, mainly, in the dark phase. In KRAS-LKB1 models, the locomotion reduction appeared more pronounced than in KRAS models.DiscussionsTransgenic animal models represent a fundamental tool to study the biology of cancers and the development of new therapies. The tumors induced in these models harbor the same genotypical and phenotypical characteristics as their human counterparts. DVC methods permit a home cage monitoring system useful for tracking animal behavior continuously 24 hours a day, 7 days a week. DVC system could determine disease progression by monitoring a single animal activity in a cage and also using group-housed animals. For these reasons, the DVC system could play a crucial role in identifying diseases at early stages and in testing new therapeutic approaches with a higher likelihood of efficacy.

IntroductionOur understanding of laboratory animal behavior and the implications of husbandry activities on their wellbeing remains incomplete. This is especially relevant with an aging colony as their activity patterns may change as they mature. Home Cage Monitoring (HCM) provides valuable insights into mouse activity within the animal's own environment and can shed light on acclimatization periods and responses to husbandry activities such as cage changing. The aim of this study was to monitor and explore changes in the activity and rest disturbance (RDI) patterns of an aging colony of male and female C57/BL6 mice.MethodsThe mice were housed in the Digitally Ventilated Cage® system, for up to 18 months of age. Data was then downloaded to investigate how the activity patterns and RDI of the mice changed over time. Habituation, aging and cage change assessments were conducted using linear mixed models, while cage separation and stereotypic behavior investigations were conducted by visual inspection of the data.ResultsAs expected during the study, mice were less active during the light phase compared to the dark phase. However, on arrival mice displayed heightened activity and RDI during the light phase and reduced activity and RDI during the dark phase, taking several days to adjust to baseline “acclimatized” patterns. With age, overall activity significantly decreased from 5 months until 14 months of age, after which it increased back toward baseline levels. We also observed activity spikes during our monitoring of this colony. Prolonged housing can lead to alarming stereotypic behaviors in animals. Cages of mice flagged for potential stereotypy displayed sustained activity spikes in the light and dark phases. Spikes in activity during the dark phase were much more pronounced than in the light phase. Cage changing led to an increase in the light phase activity and RDI compared to the previous day, with no observed difference in the dark phase post-cage change. This effect remained consistent as the animals aged.DiscussionThis study explores changes in the activity patterns of an aging colony of male and female C57/BL6 mice housed in the Digitally Ventilated Cage® system. We identified distinct aging phases concerning activity and RDI differences and a potential new welfare application for the DVC®, specifically for early detection of stereotypy. In conclusion, the adoption of HCM systems should be considered for long-term animal housing from both a welfare and behavioral perspective.

Locomotor activity can serve as a readout to identify discomfort and pain. Therefore, monitoring locomotor activity following interventions that induce potential discomfort may serve as a reliable method for evaluating animal health, complementing conventional methods such as body weight measurement. In this study, we used the digital ventilated cage (DVC®) system for the assessment of circadian locomotor activity, in addition to body weight monitoring, following intracranial stereotaxic surgery in an Alzheimer’s disease mouse model (C57BL/6J/APPswe/PSEN1dE9). Stereotaxic surgery did not affect the organization of circadian locomotor activity of both 7–8-week-old and 19–21-week-old mice. However, we observed that both young and old mice exhibited a significant decrease in activity during the dark phase. Also, our study shows that changes in locomotor activity exhibit higher sensitivity in detecting alterations indicative of animal health compared to measuring body weight. In contrast to 7–8-week-old mice, where we observed no genotypic differences in locomotor activity, 19–21-week-old APP/PS1 mice showed increased locomotor activity compared to wild-type mice. Furthermore, our analyses revealed that a subset of the 7–8-week-old mice showed increased locomotor activity during the initial peak of the dark phase. One mouse experienced sudden death early in life, possibly due to epileptic seizures. Altogether, our findings affirm continuous activity measurements as used in the DVC® as a highly valuable objective method for post-surgical welfare monitoring. Its discerning capacity not only facilitates circadian locomotor rhythm assessment but also enables the identification of individual aberrant activity patterns, possibly indicative of epileptic seizures.

Abstract Background Traditionally, in biomedical animal research, laboratory rodents are individually examined in test apparatuses outside of their home cages at selected time points. However, the outcome of such tests can be influenced by various factors and valuable information may be missed when the animals are only monitored for short periods. These issues can be overcome by longitudinally monitoring mice and rats in their home cages. To shed light on the development of home cage monitoring (HCM) and the current state-of-the-art, a systematic review was carried out on 521 publications retrieved through PubMed and Web of Science. Results Both the absolute (~ × 26) and relative (~ × 7) number of HCM-related publications increased from 1974 to 2020. There was a clear bias towards males and individually housed animals, but during the past decade (2011–2020), an increasing number of studies used both sexes and group housing. In most studies, animals were kept for short (up to 4 weeks) time periods in the HCM systems; intermediate time periods (4–12 weeks) increased in frequency in the years between 2011 and 2020. Before the 2000s, HCM techniques were predominantly applied for less than 12 h, while 24-h measurements have been more frequent since the 2000s. The systematic review demonstrated that manual monitoring is decreasing in relation to automatic techniques but still relevant. Until (and including) the 1990s, most techniques were applied manually but have been progressively replaced by automation since the 2000s. Independent of the year of publication, the main behavioral parameters measured were locomotor activity, feeding, and social behaviors; the main physiological parameters were heart rate and electrocardiography. External appearance-related parameters were rarely examined in the home cages. Due to technological progress and application of artificial intelligence, more refined and detailed behavioral parameters have been investigated in the home cage more recently. Conclusions Over the period covered in this study, techniques for HCM of mice and rats have improved considerably. This development is ongoing and further progress as well as validation of HCM systems will extend the applications to allow for continuous, longitudinal, non-invasive monitoring of an increasing range of parameters in group-housed small rodents in their home cages.

Undisturbed home cage recording of mouse activity and behavior has received increasing attention in recent years. In parallel, several technologies have been developed in a bid to automate data collection and interpretation. Thanks to these expanding technologies, massive datasets can be recorded and saved in the long term, providing a wealth of information concerning animal wellbeing, clinical status, baseline activity, and subsequent deviations in case of experimental interventions. Such large datasets can also serve as a long-term reservoir of scientific data that can be reanalyzed and repurposed upon need. In this review, we present how the impact of Big Data deriving from home cage monitoring (HCM) data acquisition, particularly through Digital Ventilated Cages (DVCs), can support the application of the 3Rs by enhancing Refinement, Reduction, and even Replacement of research in animals.

The IntelliCage allows automated testing of cognitive abilities of mice in a social home cage environment without handling by human experimenters. Restricted water access in combination with protocols in which only correct responses give access to water is a reliable learning motivator for hippocampus-dependent tasks assessing spatial memory and executive function. However, water restriction may negatively impact on animal welfare, especially in poor learners. To better comply with the 3R principles, we previously tested protocols in which water was freely available but additional access to sweetened water could be obtained by learning a task rule. While this purely appetitive motivation worked for simple tasks, too many mice lost interest in the sweet reward during more difficult hippocampus-dependent tasks. In the present study, we tested a battery of increasingly difficult spatial tasks in which water was still available without learning the task rule, but rendered less attractive either by adding bitter tasting quinine or by increasing the amount of work to obtain it. As in previous protocols, learning of the task rule provided access to water sweetened with saccharin. The two approaches of dual motivation were tested in two cohorts of female C57BL/6 N mice. Compared to purely appetitive motivation, both novel protocols strongly improved task engagement and increased task performance. Importantly, neither of the added disincentives had an adverse impact on liquid consumption, health status or body weight of the animals. Our results show that it is possible to refine test protocols in the IntelliCage so that they challenge cognitive functions without restricting access to water. [ABSTRACT FROM AUTHOR]

Background: Traditionally, in biomedical animal research, laboratory rodents are individually examined in test apparatuses outside of their home cages at selected time points. However, the outcome of such tests can be influenced by various factors and valuable information may be missed when the animals are only monitored for short periods. These issues can be overcome by longitudinally monitoring mice and rats in their home cages. To shed light on the development of home cage monitoring (HCM) and the current state-of-the-art, a systematic review was carried out on 521 publications retrieved through PubMed and Web of Science. Results: Both the absolute (~ × 26) and relative (~ × 7) number of HCM-related publications increased from 1974 to 2020. There was a clear bias towards males and individually housed animals, but during the past decade (2011–2020), an increasing number of studies used both sexes and group housing. In most studies, animals were kept for short (up to 4 weeks) time periods in the HCM systems; intermediate time periods (4–12 weeks) increased in frequency in the years between 2011 and 2020. Before the 2000s, HCM techniques were predominantly applied for less than 12 h, while 24-h measurements have been more frequent since the 2000s. The systematic review demonstrated that manual monitoring is decreasing in relation to automatic techniques but still relevant. Until (and including) the 1990s, most techniques were applied manually but have been progressively replaced by automation since the 2000s. Independent of the year of publication, the main behavioral parameters measured were locomotor activity, feeding, and social behaviors; the main physiological parameters were heart rate and electrocardiography. External appearance-related parameters were rarely examined in the home cages. Due to technological progress and application of artificial intelligence, more refined and detailed behavioral parameters have been investigated in the home cage more recently. Conclusions: Over the period covered in this study, techniques for HCM of mice and rats have improved considerably. This development is ongoing and further progress as well as validation of HCM systems will extend the applications to allow for continuous, longitudinal, non-invasive monitoring of an increasing range of parameters in group-housed small rodents in their home cages. [ABSTRACT FROM AUTHOR]

Rationale: Non-invasive home cage monitoring is emerging as a valuable tool to assess the effects of experimental interventions on mouse behaviour. A field in which these techniques may prove useful is the study of repeated selective serotonin reuptake inhibitor (SSRI) treatment and discontinuation. SSRI discontinuation syndrome is an under-researched condition that includes the emergence of sleep disturbances following treatment cessation. Objectives: We used passive infrared (PIR) monitoring to investigate changes in activity, sleep, and circadian rhythms during repeated treatment with the SSRI paroxetine and its discontinuation in mice. Methods: Male mice received paroxetine (10 mg/kg/day, s.c.) for 12 days, then were swapped to saline injections for a 13 day discontinuation period and compared to mice that received saline injections throughout. Mice were continuously tracked using the Continuous Open Mouse Phenotyping of Activity and Sleep Status (COMPASS) system. Results: Repeated paroxetine treatment reduced activity and increased behaviourally-defined sleep in the dark phase. These effects recovered to saline-control levels within 24 h of paroxetine cessation, yet there was also evidence of a lengthening of sleep bouts in the dark phase for up to a week following discontinuation. Conclusions: This study provides the first example of how continuous non-invasive home cage monitoring can be used to detect objective behavioural changes in activity and sleep during and after drug treatment in mice. These data suggest that effects of paroxetine administration reversed soon after its discontinuation but identified an emergent change in sleep bout duration, which could be used as a biomarker in future preclinical studies to prevent or minimise SSRI discontinuation symptoms. [ABSTRACT FROM AUTHOR]

There is still limited mechanistic insight into how the interaction of individuals with their environment results in the emergence of individuality in behavior and brain structure. Nevertheless, the idea that personal activity shapes the brain is implicit in strategies for healthy cognitive aging as well as in the idea that individuality is reflected in the brain's connectome. We have shown that even isogenic mice kept in a shared enriched environment (ENR) developed divergent and stable social and exploratory trajectories. As these trajectories—measured as roaming entropy (RE)—positively correlated with adult hippocampal neurogenesis, we hypothesized that a feedback between behavioral activity and adult hippocampal neurogenesis might be a causal factor in brain individualization. We used cyclin D2 knockout mice with constitutively extremely low levels of adult hippocampal neurogenesis and their wild‐type littermates. We housed them for 3 months in a novel ENR paradigm, consisting of 70 connected cages equipped with radio frequency identification antennae for longitudinal tracking. Cognitive performance was evaluated in the Morris Water Maze task (MWM). With immunohistochemistry we confirmed that adult neurogenesis correlated with RE in both genotypes and that D2 knockout mice had the expected impaired performance in the reversal phase of the MWM. But whereas the wild‐type animals developed stable exploratory trajectories with increasing variance, correlating with adult neurogenesis, this individualizing phenotype was absent in D2 knockout mice. Here the behaviors started out more random and revealed less habituation and low variance. Together, these findings suggest that adult neurogenesis contributes to experience‐dependent brain individualization. [ABSTRACT FROM AUTHOR]

Myotonic dystrophy type 1 (DM1) is a dominantly inherited neuromuscular disease caused by the abnormal expansion of CTG-repeats in the 30- untranslated region of the Dystrophia Myotonica Protein Kinase (DMPK) gene, characterized by multisystemic symptoms including muscle weakness, myotonia, cardio-respiratory problems, hypersomnia, cognitive dysfunction and behavioral abnormalities. Sleep-related disturbances are among the most reported symptoms that negatively affect the quality of life of patients and that are present in early and adult-onset forms of the disease. DMSXL mice carry a mutated human DMPK transgene containing >1,000 CTGrepeats, modeling an early onset, severe form of DM1. They exhibit a pathologic neuromuscular phenotype and also synaptic dysfunction resulting in neurological and behavioral deficits similar to those observed in patients. Additionally, they are underweight with a very high mortality within the first month after birth presenting several welfare issues. To specifically explore sleep/rest-related behaviors of this frail DM1 mouse model we used an automated home cage-based system that allows 24/7 monitoring of their activity non-invasively. We tested male and female DMSXL mice and their wild-type (WT) littermates in Digital Ventilated Cages (DVCR) assessing activity and rest parameters on day and night for 5 weeks. We demonstrated that DMSXL mice show reduced activity and regularity disruption index (RDI), higher percentage of zero activity per each hour and longer periods of rest during the active phase compared to WT. This novel rest-related phenotype in DMSXL mice, assessed unobtrusively, could be valuable to further explore mechanisms and potential therapeutic interventions to alleviate the very common symptom of excessive daytime sleepiness in DM1 patients. [ABSTRACT FROM AUTHOR]

The key goal in biomedical research is a better understanding of disease aetiologies, which ideally results in strategies and recommendations for the prevention of diseases before they arise, and in the development of effective therapies. However, many concerns have been expressed about the reproducibility and the translational validity of preclinical research in animal models to inform clinical trials in humans. It has been proposed that improving internal, external and construct validity of animal studies will lead to improved translatability. Automated behaviour monitoring in the animal's home cage, which allows for longitudinal assessment of individual trajectories over sufficiently long intervals for (chronic) drug treatment or phenotype progression, is a promising solution to these problems. [ABSTRACT FROM AUTHOR]

Keywords: home cage monitoring; cognitive function; locomotor activity; animal welfare; human disease models; early disease symptom detection EN home cage monitoring cognitive function locomotor activity animal welfare human disease models early disease symptom detection 1 5 5 06/27/22 20220623 NES 220623 Novel and emerging technologies, such as home cage monitoring (HCM) systems, permit 24/7 collection of behavioral data under undisturbed conditions. Cognitive function, locomotor activity, animal welfare, human disease models, early disease symptom detection, home cage monitoring Interestingly, animals in red tinted cages, which block most visible light, did not display such cage variation in activity between cage locations. These results demonstrate the potential of how automated HCM can aid both, animal welfare assessment and viral infection-induced symptom detection of disease in mice similar as in humans (Gaburro, [4]; Kaufmann et al., [5]). [Extracted from the article]

Some of our breeding programs include the use of Prm1 male Homozygous mice which are naturally sterile. This removes the need to use vasectomized males to induce pseudopregnancy in female mice. These males can be kept for up to 9 months and are housed with a companion female. During the timed mating period the companion female is replaced with a new female. This procedure can occur at regular intervals causing a significant increase in cage activity; one of our objectives was to determine whether this was as a result of timed mating. We wanted to investigate the disruption caused to mice during the day of the swap and how long it would take for the cage activity to return to pre-replacement baseline levels. We hypothesized that this impact would be reflected as a significant increase in cage activity, which in itself may not be a result of a negative experience but the potential of repeated disruption to their activity pattern should be considered. We used a well-known home-cage monitoring system to assess changes to the activity pattern in cages when a companion female is replaced. Data from our initial study showed that in the 2-h period after the female is replaced there is a significant increase in cage activity compared to the same time frame on the previous day. In the subsequent study, where no cage change occurred, an increase in activity was also observed when females were replaced; this returned to baseline after approximately 4 h. Prolonged activity during the rest period of mice (over 2 h) could lead to them being fatigued during their active period; therefore, as a refinement we propose that timed matings be performed later in the day, at a time when the animals are active.

Some of our breeding programs include the use of Prm1 male Homozygous mice which are naturally sterile. This removes the need to use vasectomized males to induce pseudopregnancy in female mice. These males can be kept for up to 9 months and are housed with a companion female. During the timed mating period the companion female is replaced with a new female. This procedure can occur at regular intervals causing a significant increase in cage activity; one of our objectives was to determine whether this was as a result of timed mating. We wanted to investigate the disruption caused to mice during the day of the swap and how long it would take for the cage activity to return to pre-replacement baseline levels. We hypothesized that this impact would be reflected as a significant increase in cage activity, which in itself may not be a result of a negative experience but the potential of repeated disruption to their activity pattern should be considered. We used a well-known home-cage monitoring system to assess changes to the activity pattern in cages when a companion female is replaced. Data from our initial study showed that in the 2-h period after the female is replaced there is a significant increase in cage activity compared to the same time frame on the previous day. In the subsequent study, where no cage change occurred, an increase in activity was also observed when females were replaced; this returned to baseline after approximately 4 h. Prolonged activity during the rest period of mice (over 2 h) could lead to them being fatigued during their active period; therefore, as a refinement we propose that timed matings be performed later in the day, at a time when the animals are active. [ABSTRACT FROM AUTHOR]

Amyotrophic Lateral Sclerosis (ALS) is a devastating neurodegenerative disease that affects both central and peripheral nervous system, leading to the degeneration of motor neurons, which eventually results in muscle atrophy, paralysis, and death. Sleep disturbances are common in patients with ALS, leading to even further deteriorated quality of life. Investigating methods to potentially assess sleep and rest disturbances in animal models of ALS is thus of crucial interest. We used an automated home cage monitoring system (DVC®) to capture irregular activity patterns that can potentially be associated with sleep and rest disturbances and thus to the progression of ALS in the SOD1G93A mouse model. DVC® enables non-intrusive 24/7 long term animal activity monitoring, which we assessed together with body weight decline and neuromuscular function deterioration measured by grid hanging and grip strength tests in male and female mice from 7 until 24 weeks of age. We show that as the ALS progresses over time in SOD1G93A mice, activity patterns start becoming irregular, especially during day time, with frequent activity bouts that are neither observed in control mice nor in SOD1G93A at a younger age. The increasing irregularities of activity pattern are quantitatively captured by designing a novel digital biomarker, referred to as Regularity Disruption Index (RDI). We show that RDI is a robust measure capable of detecting home cage activity patterns that could be related to rest/sleep-related disturbances during the disease progression. Moreover, the RDI rise during the early symptomatic stage parallels grid hanging and body weight decline. The non-intrusive long-term continuous monitoring of animal activity enabled by DVC® has been instrumental in discovering novel activity patterns potentially correlated, once validated, with sleep and rest disturbances in the SOD1G93A mouse model of the ALS disease.

Amyotrophic Lateral Sclerosis (ALS) is a devastating neurodegenerative disease that affects both central and peripheral nervous system, leading to the degeneration of motor neurons, which eventually results in muscle atrophy, paralysis, and death. Sleep disturbances are common in patients with ALS, leading to even further deteriorated quality of life. Investigating methods to potentially assess sleep and rest disturbances in animal models of ALS is thus of crucial interest. We used an automated home cage monitoring system (DVC®) to capture irregular activity patterns that can potentially be associated with sleep and rest disturbances and thus to the progression of ALS in the SOD1G93A mouse model. DVC® enables non-intrusive 24/7 long term animal activity monitoring, which we assessed together with body weight decline and neuromuscular function deterioration measured by grid hanging and grip strength tests in male and female mice from 7 until 24 weeks of age. We show that as the ALS progresses over time in SOD1G93A mice, activity patterns start becoming irregular, especially during day time, with frequent activity bouts that are neither observed in control mice nor in SOD1G93A at a younger age. The increasing irregularities of activity pattern are quantitatively captured by designing a novel digital biomarker, referred to as Regularity Disruption Index (RDI). We show that RDI is a robust measure capable of detecting home cage activity patterns that could be related to rest/sleep-related disturbances during the disease progression. Moreover, the RDI rise during the early symptomatic stage parallels grid hanging and body weight decline. The non-intrusive long-term continuous monitoring of animal activity enabled by DVC® has been instrumental in discovering novel activity patterns potentially correlated, once validated, with sleep and rest disturbances in the SOD1G93A mouse model of the ALS disease. [ABSTRACT FROM AUTHOR]

Highlights • Translational research requires the use of relevant preclinical pain models and assessment tools. • Clinically relevant behavioral tests should focus on restoring function by reducing pathological pain in the absence of disruptive side effects. • Grimace, Operant Behavior, Wheel Running, Burrowing, Nesting, Home Cage Monitoring, Gait Analysis, CPP and CPA are described. • Analysis of the advantages and limitations of these tests will help researchers identify appropriate tests for their particular goal. Abstract The primary objective of preclinical pain research is to improve the treatment of pain. Decades of research using pain-evoked tests has revealed much about mechanisms but failed to deliver new treatments. Evoked pain-tests are often limited because they ignore spontaneous pain and motor or disruptive side effects confound interpretation of results. New tests have been developed to focus more closely on clinical goals such as reducing pathological pain and restoring function. The objective of this review is to describe and discuss several of these tests. We focus on: Grimace Scale, Operant Behavior, Wheel Running, Burrowing, Nesting, Home Cage Monitoring, Gait Analysis and Conditioned Place Preference/ Aversion. A brief description of each method is presented along with an analysis of the advantages and limitations. The pros and cons of each test will help researchers identify the assessment tool most appropriate to meet their particular objective to assess pain in rodents. These tests provide another tool to unravel the mechanisms underlying chronic pain and help overcome the translational gap in drug development. [ABSTRACT FROM AUTHOR]

Background: There is an urgent need to develop and incorporate novel behavioral tests in classically used preclinical pain models. Most rodent studies are based upon stimulus-evoked hindpaw measurements even though chronic pain is usually a day and night experience. Chronic pain is indeed a debilitating condition that influences the sociability and the ability for voluntary tasks, but the relevant behavioral readouts for these aspects are mostly under-represented in the literature. Moreover, we lack standardization in most behavioral paradigms to guarantee reproducibility and ensure adequate discussion between different studies. This concerns not only the combination, application, and duration of particular behavioral tasks but also the effects of different housing conditions implicating social isolation. Results: Our aim was to thoroughly characterize the classically used spared nerve injury model for 12 weeks following surgery. We used a portfolio of classical stimulus-evoked response measurements, detailed gait analysis with two different measuring systems (Dynamic weight bearing (DWB) system and CatWalk), as well as observer-independent voluntary wheel running and home cage monitoring (Laboras system). Additionally, we analyzed the effects of social isolation in all behavioral tasks. We found that evoked hypersensitivity temporally matched changes in static gait parameters, whereas some dynamic gait parameters were changed in a time-dependent manner. Interestingly, voluntary wheel running behavior was not affected in spared nerve injury mice but by social isolation. Besides a reduced climbing activity, spared nerve injury mice did not showed tremendous alterations in the home cage activity. Conclusion: This is the first longitudinal study providing detailed insights into various voluntary behavioral parameters related to pain and highlights the importance of social environment on spontaneous non-evoked behaviors in a mouse model of chronic neuropathy. Our results provide fundamental considerations for future experimental planning and discussion of pain-related behavioral changes. [ABSTRACT FROM AUTHOR]

Background: Physical activity in Amyotrophic Lateral Sclerosis (ALS) plays a controversial role. In some epidemiological studies, both recreational or professional sport exercise has been associated to an increased risk for ALS but the mechanisms underlying the effects of exercise have not been fully elucidated in either patients or animal models., Methods: To better reproduce the influence of this environmental factor in the pathogenesis of ALS, we exposed SOD1 G93A low-copy male mice to multiple exercise sessions at asymptomatic and pre-symptomatic disease stages in an automated home-cage running-wheel system for about 3 months., Results: Repeated voluntary running negatively influenced disease progression by anticipating disease onset, impairing neuromuscular transmission, worsening neuromuscular decline, and exacerbating muscle atrophy. Muscle fibers and neuromuscular junctions (NMJ) as well as key molecular players of the nerve-muscle circuit were similarly affected., Conclusion: It thus appears that excessive physical activity can be detrimental in predisposed individuals and these findings could model the increased risk of developing ALS in predisposed and specific professional athletes., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Simple Summary: Different forms of environmental enrichment are used to increase the wellbeing of laboratory animals. These forms include extending the available cage space, housing a large group of animals within the same unit and adding stimulating physical objects. The semi naturalistic environment (SNE) used in this study implements all of these enhancements. However, there is debate as to whether such variation in housing standards increases the variability of experimental data. Indeed, it has been shown that mice living in the SNE developed individual differences in activity and behavioral parameters. Here, we investigated whether housing in the SNE enhances individual differences in aged animals and whether these differences are reflected in certain physiological parameters. These aspects were considered to assess the suitability of the SNE as a reference system in future studies. We found that the individual-level activity patterns of the animals stabilized during the housing period in the SNE. These behavioral characteristics did not correlate with the measured physiological parameters. Considering the variance of the measured data, which is comparable to the literature, the SNE seems to be a suitable system for studies comparing different housing systems in terms of animal welfare. Despite tremendous efforts at standardization, the results of scientific studies can vary greatly, especially when considering animal research. It is important to emphasize that consistent different personality-like traits emerge and accumulate over time in laboratory mice despite genetic and environmental standardization. To understand to what extent variability can unfold over time, we conducted a long-term study using inbred mice living in an exceptionally complex environment comprising an area of 4.6 m2 spread over five levels. In this semi-naturalistic environment (SNE) the activity and spatial distribution of 20 female C57Bl/6J was recorded by radio-frequency identification (RFID). All individuals were monitored from an age of 11 months to 22 months and their individual pattern of spatial movement in time is described as roaming entropy. Overall, we detected an increase of diversification in roaming behavior over time with stabilizing activity patterns at the individual level. However, spontaneous behavior of the animals as well as physiological parameters did not correlate with cumulative roaming entropy. Moreover, the amount of variability did not exceed the literature data derived from mice living in restricted conventional laboratory conditions. We conclude that even taking quantum leaps towards improving animal welfare does not inevitably mean a setback in terms of data quality. [ABSTRACT FROM AUTHOR]