Mice with endogenous TDP‐43 mutations exhibit gain of splicing function and characteristics of amyotrophic lateral sclerosis

TDP‐43 (encoded by the gene TARDBP) is an RNA binding protein central to the pathogenesis of amyotrophic lateral sclerosis (ALS). However, how TARDBP mutations trigger pathogenesis remains unknown. Here, we use novel mouse mutants carrying point mutations in endogenous Tardbp to dissect TDP‐43 function at physiological levels both in vitro and in vivo. Interestingly, we find that mutations within the C‐terminal domain of TDP‐43 lead to a gain of splicing function. Using two different strains, we are able to separate TDP‐43 loss‐ and gain‐of‐function effects. TDP‐43 gain‐of‐function effects in these mice reveal a novel category of splicing events controlled by TDP‐43, referred to as “skiptic” exons, in which skipping of constitutive exons causes changes in gene expression. In vivo, this gain‐of‐function mutation in endogenous Tardbp causes an adult‐onset neuromuscular phenotype accompanied by motor neuron loss and neurodegenerative changes. Furthermore, we have validated the splicing gain‐of‐function and skiptic exons in ALS patient‐derived cells. Our findings provide a novel pathogenic mechanism and highlight how TDP‐43 gain of function and loss of function affect RNA processing differently, suggesting they may act at different disease stages.     

Comparison of sensory structures on the antenna of different species of Philopotamidae (Insecta: Trichoptera)

Structure and distribution of sensilla were studied in sixteen species of the caddisfly family Philopotamidae. Their antennae bear numerous curved trichoid and pseudoplacoid sensilla and fewer coronal, styloconic and chaetoid sensilla on the flagellar segments. The most numerous pseudoplacoid sensilla have non-specific localization. The curved trichoid sensilla form clusters ventrally on each antennal segment. Sensilla belonging to coronal, styloconic and chaetoid types have specific positions. Long grooved trichoid sensilla are located nonspecifically in all the studied species. The average number of sensilla per segment decreases from the proximal to distal part of the flagellum. Scapus and pedicellum are devoid of most types of sensilla, however, they bear the Böhm bristles and long trichoid sensilla. A positive correlation between antenna dimensions and its cuticular structures is found.     

Spatial Patterns of Primate Electrocutions in Diani,Kenya

Electrocution from power infrastructure threatens many primate species, yet knowledge of effective evidence-based mitigation strategies is limited. Mitigation planning requires an understanding of the spatial distribution of electrocutions to prioritize high-risk areas. In Diani, a coastal Kenyan town, electrocution is an important cause of death for five primate species. In this study we aim to describe the spatial patterns of electrocutions and electric shock incidents (collectively referred to as electrocutions hereafter) and identify electrocution hotspots to guide an effective primate conservation approach in Diani. Colobus Conservation, a not-for-profit organization, has recorded electrocutions and annual primate census data since 1998. We georeferenced 329 electrocution data points and analyzed them using QGIS. We identified and compared hotspots across species, seasons, and time using kernel density estimation and Getis-Ord-Gi*. We employed spatial regression models to test whether primate population density and power line density predicted the location of electrocution hotspots. Electrocutions occurred in hotspots that showed little variation in location between species and seasons. The limited variation in hotspot location over time likely occurred as a result of new building development in Diani and variability in primate detection rates by community members. Primate density and power line density were significant predictors of electrocution density for Angolan black-and-white colobus (Colobus angolensis palliatus) and Sykes monkeys (Cercopithecus mitis albogularis), but the relationship was weak, suggesting the presence of additional risk factors. This study provides a framework for systematic spatial prioritization of power lines that can be used to reduce primate electrocutions in Diani, and can be adopted in other areas of the world where primates are at risk from electrocution.     

Laminin α1 Chain Synthesis in the Mouse Developing Lung: Requirement for Epithelial–Mesenchymal Contact and Possible Role in Bronchial Smooth muscle Development

Laminins, the main components of basement membranes, are heterotrimers consisting of α, β, and γ polypeptide chains linked together by disulfide bonds. Laminins-1 and -2 are both composed of β1 and γ1 chains and differ from each other on their α chain, which is α1 and α2 for laminin-1 and -2, respectively. The present study shows that whereas laminins-1 and -2 are synthesized in the mouse developing lung and in epithelial–mesenchymal cocultures derived from it, epithelial and mesenchymal monocultures lose their ability to synthesize the laminin α1 chain. Synthesis of laminin α1 chain however returns upon re-establishment of epithelial–mesenchymal contact. Cell–cell contact is critical, since laminin α1 chain is not detected in monocultures exposed to coculture-conditioned medium or in epithelial–mesenchymal cocultures in which heterotypic cell–cell contact is prevented by an interposing filter. Immunohistochemical studies on cocultures treated with brefeldin A, an inhibitor of protein secretion, indicated both epithelial and mesenchymal cells synthesize laminin α1 chain upon heterotypic cell– cell contact. In a set of functional studies, embryonic lung explants were cultured in the presence of monoclonal antibodies to laminin α1, α2, and β/γ chains. Lung explants exposed to monoclonal antibodies to laminin α1 chain exhibited alterations in peribronchial cell shape and decreased smooth muscle development, as indicated by low levels of smooth muscle α actin and desmin. Taken together, our studies suggest that laminin α1 chain synthesis is regulated by epithelial–mesenchymal interaction and may play a role in airway smooth muscle development.     

GIANT KELP (MACROCYSTIS PYRIFERA, PHAEOPHYCEAE) RECRUITMENT NEAR ITS SOUTHERN LIMIT IN BAJA CALIFORNIA AFTER MASS DISAPPEARANCE DURING ENSO 1997–1998

During the ENSO event of 1997–1998, density and population structure were evaluated in a Macrocystis pyrifera forest located in Bahía Tortugas, Baja California, Mexico, near the southern limit of the species' distribution in the Northern Hemisphere. Observations in Bahía Tortugas were made quarterly from January 1997 to September 1998 using SCUBA diving surveys. No macroscopic plants were found in the Bahía Tortugas area from October 1997 to April 1998, a local absence of at least 7 months. Aerial surveys further suggest regional disappearance along most of the Baja California coast during the event. Unexpectedly, plants were found in Bahía Tortugas again in July 1998, in spite of the widespread disappearance of the species less than a year earlier. Long-distance spore dispersal was an unlikely cause of the recruitment because: 1) the nearest spore source was more than 100 km away; 2) recruitment appeared to be simultaneous at many sites and occurred rapidly after the cessation of the ENSO event; and 3) the recruits occurred in the same areas as before disappearance. We suggest that a microscopic stage that was not visible during dive surveys survived the stressful conditions of ENSO and caused the recruitment event, supporting the hypothesis that a bank of microscopic forms can survive conditions stressful to macroscopic algae.     

Laminin α4 and Integrin α6 Are Upregulated in Regenerating dy/dy Skeletal Muscle: Comparative Expression of Laminin and Integrin Isoforms in Muscles Regenerating after Crush Injury

The expression of laminin isoforms and laminin-binding integrin receptors known to occur in muscle was investigated during myogenic regeneration after crush injury. Comparisons were made between dystrophic 129ReJ dy/dy mice, which have reduced laminin α2 expression, and their normal littermates. The overall histological pattern of regeneration after crush injury was similar in dy/dy and control muscle, but proceeded faster in dy/dy mice. In vitro studies revealed a greater yield of mononuclear cells extracted from dy/dy muscle and a reduced proportion of desmin-positive cells upon in vitro cultivation, reflecting the presence of inflammatory cells and “preactivated” myoblasts due to ongoing regenerative processes within the endogenous dystrophic lesions. Laminin α1 was not detectable in skeletal muscle. Laminin α2 was present in basement membranes of mature myofibers and newly formed myotubes in control and dy/dy muscles, albeit weaker in dy/dy. Laminin α2-negative myogenic cells were detected in dy/dy and control muscle, suggesting the involvement of other laminin α chains in early myogenic differentiation, such as laminin α4 and α5 which were both transiently expressed in basement membranes of newly formed myotubes of dy/dy and control mice. Integrin β1 was expressed on endothelial cells, muscle fibers, and peripheral nerves in uninjured muscle and broadened after crush injury to the interstitium where it occurred on myogenic and nonmyogenic cells. Integrin α3 was not expressed in uninjured or regenerating muscle, while integrin α6 was expressed mainly on endothelial cells and peripheral nerves in uninjured muscle. Upon crush injury integrin α6 increased in the interstitium mainly on nonmyogenic cells, including infiltrating leukocytes, endothelial cells, and fibroblasts. In dy/dy muscle, integrin α6 occurred on some newly formed myotubes. Integrin α7 was expressed on muscle fibers at the myotendinous junction and showed weak and irregular expression on muscle fibers. After crush injury, integrin α7 expression extended to the newly formed myotubes and some myoblasts. However, many myoblasts and newly formed myotubes were integrin α7 negative. No marked difference was observed in integrin α7 expression between dy/dy and control muscle, either uninjured or after crush injury. Only laminin α4 and integrin α6 expression patterns were notably different between dy/dy and control muscle. Expression of both molecules was more extensive in dy/dy muscle, especially in the interstitium of regenerating areas and on newly formed myotubes. In view of the faster myogenic regeneration observed in dy/dy mice, the data suggest that laminin α4 and integrin α6 support myogenic regeneration. However, whether these accelerated myogenic effects are a direct consequence of the reduced laminin α2 expression in dy/dy mice, or an accentuation of the ongoing regenerative events in focal lesions in the muscle, requires further investigation.