Dystrophin deficiency leads to disturbance of LAMP1-vesicle-associated protein secretion

Duchenne muscular dystrophy results from loss of the protein dystrophin, which links the intracellular cytoskeletal network with the extracellular matrix, but deficiency in this function does not fully explain the onset or progression of the disease. While some intracellular events involved in the degeneration of dystrophin-deficient muscle fibers have been well characterized, changes in their secretory profile are undescribed. To analyze the secretome profile of mdx myotubes independently of myonecrosis, we labeled the proteins of mdx and wild-type myotubes with stable isotope-labeled amino acids (SILAC), finding marked enrichment of vesicular markers in the mdx secretome. These included the lysosomal-associated membrane protein, LAMP1, that co-localized in vesicles with an over-secreted cytoskeletal protein, myosin light chain 1. These LAMP1/MLC1-3-positive vesicles accumulated in the cytosol of mdx myotubes and were secreted into the culture medium in a range of abnormal densities. Restitution of dystrophin expression, by exon skipping, to some 30 % of the control value, partially normalized the secretome profile and the excess LAMP1 accumulation. Together, our results suggest that a lack of dystrophin leads to a general dysregulation of vesicle trafficking. We hypothesize that disturbance of the export of proteins through vesicles occurs before, and then concurrently with, the myonecrotic cascade and contributes chronically to the pathophysiology of DMD, thereby presenting us with a range of new potential therapeutic targets.     

Natural enemy assemblages on native and reseeded grasslands in southwestern Montana: a family-level analysis

We conducted a 2-year survey, using sweep sampling and family-level taxon identification, of the predatory and parasitoid insects on grassland sites in the Gallatin Valley of southwestern Montana. The 25 sites were divided into 4 habitat classes: 2 native habitat types ( Stipa comata/Bouteloua gracilis and Festuca idahoensis/Agropyron spicatum ) and 2 that had been reseeded with either crested wheatgrass ( Agropyron cristatum ) or smooth brome ( Bromus inermis ). Our major goal was to make quantitative comparisons of the abundance of insects among native and reseeded habitats. Of 51 families in 5 insect orders identified, 7 Hymenoptera (Encyrtidae, Braconidae, Ichneumonidae, Pteromalidae, Eulophidae, Scelionidae, and Torymidae), 3 Hemiptera (Lygaeidae, Nabidae, and Reduviidae), 1 Coleoptera (Coccinellidae), and 1 Diptera (Asilidae) comprised 90% of the natural enemies sampled. Ordination analyses provided no strong evidence that the 4 habitat classes contained distinct overall natural enemy communities. However, contiguous native and reseeded sites usually had relatively different overall natural enemy assemblages, suggesting that vegetation was often a more important correlate of community composition than was close spatial proximity of sites. Furthermore, several common families exhibited differential abundances across habitat classes in one or both years. For example, in 1989, Eulophidae, Pteromalidae, and Torymidae were more abundant on native Festuca/Agropyron sites, whereas Encyrtidae and Nabidae were more abundant on Festuca/Agropyron sites reseeded with Bromus inermis . Although analyses of insect assemblages classified to the family level provide somewhat limited information on functional ecological differences among habitats, they allow one to survey a broad array of taxa to identify focal groups for future conservation and land management studies.