Background and Purpose: Amyotrophic lateral sclerosis (ALS) patients exhibit dysfunctional energy metabolism and weight loss, which is negatively correlated with survival, together with neuroinflammation. However, the possible contribution of neuroinflammation to deregulations of feeding behaviour in ALS has not been studied in detail. We here investigated if microglial K_Ca3.1 is linked to hypothalamic neuroinflammation and affects feeding behaviours in ALS mouse models.Experimental Approach: hSOD1^G93A and TDP43^A315T mice were treated daily with 120 mg·kg⁻¹ of TRAM‐34 or vehicle by intraperitoneal injection from the presymptomatic until the disease onset phase. Body weight and food intake were measured weekly. The later by weighing food provided minus that left in the cage. RT‐PCR and immunofluorescence analysis were used to characterize microglia phenotype and the main populations of melanocortin neurons in the hypothalamus of hSOD1^G93A and age‐matched non‐tg mice. The cannabinoid–opioid interactions in feeding behaviour of hSOD1^G93A mice were studied using an inverse agonist and an antagonist of the cannabinoid receptor CB₁ (rimonabant) and μ‐opioid receptors (naloxone), respectively.Key Results: We found that treatment of hSOD1^G93A mice with the K_Ca3.1 inhibitor TRAM‐34 (i), attenuates the pro‐inflammatory phenotype of hypothalamic microglia, (ii) increases food intake and promotes weight gain, (iii) increases the number of healthy pro‐opiomelanocortin (POMC) neurons and (iv), changes the expression of cannabinoid receptors involved in energy homeostasis.Conclusion and Implications: Using ALS mouse models, we describe defects in the hypothalamic melanocortin system that affect appetite control. These results reveal a new regulatory role for K_Ca3.1 to counteract weight loss in ALS.

Early treatment of ischemic stroke can significantly reduce disability and mortality rates. Stem cell-derived extracellular vesicles (EVs) have shown potential as therapeutics for neurological disorders. This study explored whether intranasal administration of EVs from human bone marrow mesenchymal stem cells (BM-MSCs) enhances forelimb motor function recovery in a mouse model of motor cortex stroke and investigated their mechanism of action, focusing on neuroinflammation. C57BL/6JRj mice received EV treatment of 0.1 × 10⁹ EVs per dose per day, 48 h post-stroke and twice weekly for four weeks. EV-treated mice showed significant improvement in forelimb deficits, as evaluated using a series of motor tests. Histopathological assessments revealed reduced infarct volume and decreased astrogliosis and microglial activation in EV-treated mice. EV treatment led to changes in microglial morphology in the peri-infarct area, associated with increased anti-inflammatory cytokines interleukin (IL)-10 and IL-13 and decreased pro-inflammatory cytokines IL-1β, IL-6, and tumor necrosis factor-alpha. Reduced expression of nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome and Cleaved Caspase-1 following EV treatment supports their role in dampening inflammation. In vitro experiments using oxygen-glucose deprivation confirmed that EVs attenuated the inflammatory phenotype of microglia and reduced neuronal apoptosis. EV cargo analysis revealed neuroprotective molecules, including anti-inflammatory cytokines and brain-derived neurotrophic factor (BDNF), which may contribute to their immunomodulatory properties. These findings show that EVs mitigate post-stroke brain immune response, promoting tissue healing and recovery. Our comprehensive characterization of the effects of human BM-MSC-derived EVs, encompassing functional, tissue, cellular, and molecular aspects, underscores their therapeutic potential and supports their use in stroke treatment.

In recent years, significant advancements have been made in utilizing nanoparticles (NPs) to modulate immune responses within the central nervous system (CNS), offering new opportunities for nanotherapeutic interventions in neurological disorders. NPs can serve as carriers for immunomodulatory agents or platforms for delivering nucleic acid-based therapeutics to regulate gene expression and modulate immune responses. Several studies have demonstrated the efficacy of NP-mediated immune modulation in preclinical models of neurological diseases, including multiple sclerosis, stroke, Alzheimer’s disease, and Parkinson’s disease. While challenges remain, advancements in NPs engineering and design have led to the development of NPs using diverse strategies to overcome these challenges. The nano-bio interface with the immune system is key in the conceptualization of NPs to efficiently act as nanotherapeutics in the CNS. The biomolecular corona plays a pivotal role in dictating NPs behavior and immune recognition within the CNS, giving researchers the opportunity to optimize NPs design and surface modifications to minimize immunogenicity and enhance biocompatibility. Here, we review how NPs interact with the CNS immune system, focusing on immunosurveillance of NPs, NP-induced immune reprogramming and the impact of the biomolecular corona on NPs behavior in CNS immune responses. The integration of NPs into CNS nanotherapeutics offers promising opportunities for addressing the complex challenges of acute and chronic neurological conditions and pathologies, also in the context of preventive and rehabilitative medicine. By harnessing the nano-bio immune interface and understanding the significance of the biomolecular corona, researchers can develop targeted, safe, and effective nanotherapeutic interventions for a wide range of CNS disorders to improve treatment and rehabilitation. These advancements have the potential to revolutionize the treatment landscape of neurological diseases, offering promising solutions for improved patient care and quality of life in the future.

Duchenne muscular dystrophy (DMD) is an X-linked disease, caused by a mutant dystrophin gene, leading to muscle membrane instability, followed by muscle inflammation, infiltration of pro-inflammatory macrophages and fibrosis. The calcium-activated potassium channel type 3.1 (K_Ca3.1) plays key roles in controlling both macrophage phenotype and fibroblast proliferation, two critical contributors to muscle damage. In this work, we demonstrate that pharmacological blockade of the channel in the mdx mouse model during the early degenerative phase favors the acquisition of an anti-inflammatory phenotype by tissue macrophages and reduces collagen deposition in muscles, with a concomitant reduction of muscle damage. As already observed with other treatments, no improvement in muscle performance was observed in vivo. In conclusion, this work supports the idea that K_Ca3.1 channels play a contributing role in controlling damage-causing cells in DMD. A more complete understanding of their function could lead to the identification of novel therapeutic approaches.

Dystrophin is a cytoskeletal protein contributing to the organization of the neuromuscular junction. In Duchenne muscular dystrophy, due to dystrophin absence, the distribution of endplate acetylcholine receptors (AChRs) becomes disorganized. It is still debated whether this is due to the absence of dystrophin or to the repeated damage/regeneration cycles typical of dystrophic muscle. We addressed this controversy studying the endplate in the first 3 postnatal weeks, when muscle damage in dystrophic (mdx) mice is minimal. By synaptic and extra-synaptic patch-clamp recordings in acutely dissociated mdx and wt muscle fibers, we recorded unitary events due to openings of AChR-channels containing the γ and ε subunit. We also examined AChR distribution at the endplate by immunofluorescence assays. No differences between wt and mdx fibers were found in the γ/ε switch, nor in the AChR organization at the endplates up to 21 postnatal days. Conversely, we detected a delayed appearance and disappearance of patches with high channel opening frequency in mdx fibers. Our data emphasize that the innervation-dependent γ/ε switch and AChR organization in the endplate are not affected by the absence of dystrophin, while extra-synaptic AChR cluster formation and disassembly could be differentially regulated in mdx mice.

Understanding age-related neurophysiological changes is crucial for identifying brain aging biomarkers and developing strategies against motor and cognitive decline. To explore aging-related patterns across multiple domains, this study assessed motor and cognitive performance, functional connectivity, and synaptic organization in Young (4 months), Adult (14 months), and Old (24 months) mice. Adult mice exhibited reduced locomotor activity (−50.3%) and forelimb force (−38.3%) compared to Young mice, while Old mice showed decline in spatial (−30.4%) and recognition memory (−40.3%). Golgi–Cox staining revealed region-specific changes in spine density, including an increase in motor cortex layer II/III pyramidal neurons in Old versus Adult mice and reductions in the medial prefrontal cortex and CA1 hippocampal region. Immunofluorescence analysis indicated age-related alterations in VGLUT and VGAT expression across brain regions. Local field potential recordings revealed no significant changes in functional connectivity across age groups. Integration of behavioral and electrophysiological features using machine learning for an exploratory yielded a classification accuracy of 0.798. Although this represented only a modest and non-significant improvement over behavioral features alone, the highest pairwise discrimination was observed between Adult and Old mice (AUC = 0.861). Overall, these findings provide a multilevel descriptive analysis of brain aging, highlighting distinct behavioral and structural alterations alongside preserved functional connectivity.