Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of unknown cause. Absence of specific targets and biomarkers compromise the development of new therapeutic strategies and of innovative tools to stratify patients and assess their responses to treatment. Here, we investigate changes in neuroprotective-neuroinflammatory actions in the spinal cord of SOD1 mice, at presymptomatic and symptomatic stages to identify stage-specific biomarkers and potential targets. Results showed that in the presymptomatic stage, there are alterations in both astrocytes and microglia, which comprise decreased expression of GFAP and S100B and upregulation of GLT-1, as well as reduced expression of CD11b, M2-phenotype markers, and a set of inflammatory mediators. Reduced levels of Connexin-43, Pannexin-1, CCL21, and CX3CL1 further indicate the existence of a compromised intercellular communication. In contrast, in the symptomatic stage, increased markers of inflammation became evident, such as NF-κB/Nlrp3-inflammasome, Iba1, pro-inflammatory cytokines, and M1-polarizion markers, together with a decreased expression of M2-phenotypic markers. We also observed upregulation of the CX3CL1-CX3CR1 axis, Connexin-43, Pannexin-1, and of microRNAs (miR)-124, miR-125b, miR-146a and miR-21. Reduced motor neuron number and presence of reactive astrocytes with decreased GFAP, GLT-1, and GLAST further characterized this inflammatory stage. Interestingly, upregulation of miR-155 and downregulation of MFG-E8 appear as consistent biomarkers of both presymptomatic and symptomatic stages. We hypothesize that downregulated cellular interplay at the early stages may represent neuroprotective mechanisms against inflammation, SOD1 aggregation, and ALS onset. The present study identified a set of inflamma-miRNAs, NLRP3-inflammasome, HMGB1, CX3CL1-CX3CR1, Connexin-43, and Pannexin-1 as emerging candidates and promising pharmacological targets that may represent potential neuroprotective strategies in ALS therapy.

No abstract

The cray®sh Procambarus clarkii is an introduced species in Portugal, becoming a new available resource that has possibly changed local trophic relationships. The purpose of the present study was to understand better how a naã Ève assembly of predators reacted to a recent invader. Predation on P. clarkii by nine species of mammals (Order: Carnivora) and six species of birds (Order: Ciconiiformes) in a marsh area in Portugal, was surveyed through the collection of faeces and pellets over a 24-month period. Seasonal variation in the consumption of this prey by different predators was evaluated, and estimation made of size and weight of P. clarkii consumed by them. Procambarus clarkii was readily preyed upon by four species of mammalian carnivores and ®ve species of ciconiiform birds. The consumption of P. clarkii varied seasonally in both years with a trend between seasonal exploitation and its population structure and availability. Generally, the use of P. clarkii by all predators was more intense in spring, summer and autumn than in winter, and was directed towards larger and heavier individuals. These ®ndings result from the population dynamics presented by P. clarkii in the marsh area studied here, since larger and heavier individuals were more available in spring, summer and autumn. Procambarus clarkii is an important food resource for mammals and birds and it seems to play a key role in the trophic interactions of the riparian and terrestrial communities of the marsh.