B cell depletion modulates glial responses and enhances blood vessel integrity in a model of multiple sclerosis.

Multiple sclerosis (MS) is characterized by a compromised blood-brain barrier (BBB) resulting in central nervous system (CNS) entry of peripheral lymphocytes, including T cells and B cells. While T cells have largely been considered the main contributors to neuroinflammation in MS, the success of B cell depletion therapies suggests an important role for B cells in MS pathology. Glial cells in the CNS are essential components in both disease progression and recovery, raising the possibility that they represent targets for B cell functions. Here, we examine astrocyte and microglia responses to B cell depleting treatments in an animal model of MS, experimental autoimmune encephalomyelitis (EAE). B cell depleted EAE animals had markedly reduced disease severity and myelin damage accompanied by reduced microglia and astrocyte reactivity 20 days after symptom onset. To identify potential initial mechanisms mediating functional changes following B cell depletion, astrocyte and microglia transcriptomes were analyzed 3 days following B cell depletion. In control EAE animals, transcriptomic analysis revealed astrocytic inflammatory pathways were activated and microglial influence on neuronal function were inhibited. Following B cell depletion, initial functional recovery was associated with an activation of astrocytic pathways linked with restoration of neurovascular integrity and of microglial pathways associated with neuronal function. These studies reveal an important role for B cell depletion in influencing glial function and CNS vasculature in an animal model of MS.

Effects of Covid-19 on physical activity and eating behaviours and their mutual relationship.

The study aimed to determine the relationship between physical activity and eating behaviours among the physical therapy students of various physical therapy institutes of Rawalpindi/Islamabad during the Covid-19 pandemic. The study was conducted from September 2020 to February 2021. It was a cross sectional co-relational survey of a total of 209 physical therapy students, and the sampling technique was purposive sampling. Eating Attitude Test (EAT-26) Questionnaire and International Physical Activity Questionnaire (IPAQ) were used as the data collection tools. Healthy young students of 18-26 years of age, of both genders, were included and students with major functional limitations were excluded from the study. The results showed that 87 (41.6%) participants had normal EAT-26 scores whereas 122 (58.4%) had abnormal EAT-26 scores. IPAQ scores were categorically distributed into low, moderate, and high levels of physical activity. Eighty-seven (41.6%) had low levels of physical activity, 106 (50.7%) had moderate levels of physical activity, and 16 (7.7%) had high levels of physical activity. Spearman Rho test of correlation was applied with the p-value 0.219, and R-value of correlation was 0.08, which showed weak positive correlation between physical activity and eating behaviours. The study concluded that restricted physical activity and home confinement due to Covid-19 pandemic, did not potentially affect the physical therapy students' eating behaviours.

Isolation of Pure Astrocytes and Microglia from the Adult Mouse Spinal Cord For In Vitro Assays and Transcriptomic Studies.

Astrocytes and microglia play pivotal roles in central nervous system development, injury responses, and neurodegenerative diseases. These highly dynamic cells exhibit rapid responses to environmental changes and display significant heterogeneity in terms of morphology, transcriptional profiles, and functions. While our understanding of the functions of glial cells in health and disease has advanced substantially, there remains a need for in vitro, cell-specific analyses conducted in the context of insults or injuries to comprehensively characterize distinct cell populations. Isolating cells from the adult mouse offers several advantages over cell lines or neonatal animals, as it allows for the analysis of cells under pathological conditions and at specific time points. Furthermore, focusing on spinal cord-specific isolation, excluding brain involvement, enables research into spinal cord pathologies, including experimental autoimmune encephalomyelitis, spinal cord injury, and amyotrophic lateral sclerosis. This protocol presents an efficient method for isolating astrocytes and microglia from the adult mouse spinal cord, facilitating immediate or future analysis with potential applications in functional, molecular, or proteomic downstream studies.

Cell type specific isolation of primary astrocytes and microglia from adult mouse spinal cord.

BACKGROUND: Astrocytes and microglia are essential cellular elements of the CNS that are critical for normal development, function, and injury responses. Both cell types are highly pleiotropic and respond rapidly to environmental changes, making them challenging to characterize. One approach is to develop efficient isolation paradigms of distinct cell populations, allowing for characterization of their roles in distinct CNS regions and in pathological states. NEW METHOD: We have developed an efficient and reliable protocol for isolation of astrocytes and microglia from the adult mouse spinal cord, which can be easily manipulated for immediate or future analyses. This method involves (1) rapid tissue dissociation; (2) cell release after myelin debris removal; (3) magnetic-activated cell sorting; and (4) optional downstream molecular and functional analyses. RESULTS: High levels of viability and purity of the cells were confirmed after isolation. More importantly, characterization of cells verified their ability to proliferate and respond to external stimuli for potential use in downstream molecular and functional assays. COMPARISON WITH EXISTING METHOD(S): Long-term culture of cells isolated from neonatal animals and cell type specific isolation from the brain have been successful; however, isolation of spinal cord cells from adult mice has been challenging due to the large amount of myelin and limited size of the tissue compared to the brain. Our method allows for efficient isolation of astrocytes and microglia from spinal cord alone and includes simple modifications to allow for various downstream applications. CONCLUSIONS: This technique will be a valuable tool to better understand the functions of astrocytes and microglia in spinal cord function and pathology.