NEDD4-1 deficiency impairs satellite cell function during skeletal muscle regeneration

BACKGROUND: Satellite cells are tissue-specific stem cells primarily responsible for the regenerative capacity of skeletal muscle. Satellite cell function and maintenance are regulated by extrinsic and intrinsic mechanisms, including the ubiquitin-proteasome system, which is key for maintaining protein homeostasis. In this context, it has been shown that ubiquitin-ligase NEDD4-1 targets the transcription factor PAX7 for proteasome-dependent degradation, promoting muscle differentiation in vitro. Nonetheless, whether NEDD4-1 is required for satellite cell function in regenerating muscle remains to be determined. RESULTS: Using conditional gene ablation, we show that NEDD4-1 loss, specifically in the satellite cell population, impairs muscle regeneration resulting in a significant reduction of whole-muscle size. At the cellular level, NEDD4-1-null muscle progenitors exhibit a significant decrease in the ability to proliferate and differentiate, contributing to the formation of myofibers with reduced diameter. CONCLUSIONS: These results indicate that NEDD4-1 expression is critical for proper muscle regeneration in vivo and suggest that it may control satellite cell function at multiple levels.

Geographic Area of Collection Determines the Chemical Composition and Antimicrobial Potential of Three Extracts of Chilean Propolis.

The biological properties of chilean propolis have been described and include antibacterial, antifungal and antibiofilm activities. Propolis has a strong antimicrobial potential. Clinical experiences with synthetic antibiotics indicated the need to discover new sources of bioactive compounds associated with ethnopharmacological knowledge or natural sources such as propolis. The microscopic analysis of pollen grains from plants allows us to determine the botanical origin of the propolis samples. In Angol, sample pollen grains were obtained from fodder plants (Sorghum bicolor; Lotus sp.) and trees, such as Acacia sp., Pinus radiata, Eucalyptus sp. and Salix babylonica. Propolis from the Maule region contains pollen grains from endemic plants such as Quillaja saponaria. Finally, the sample obtained from Melipilla presented a wider variety of pollen extracted from vegetable species.Colorimetric assays performed to quantify the total polyphenols present in Chilean propolis samples established that PCP2 (Angol sample) showed high amounts of phenolics compounds, with significant statistical differences in comparison with the other samples. The main compounds identified were pinocembrin, quercetin and caffeic acid phenethyl ester (CAPE). The Angol sample showed a high content of polyphenols.Studies that determine the influence of geographical and floral variables on the chemical composition of propolis are a valuable source of information for the study of its biological properties.

Participation of the SMAD2/3 signalling pathway in the down regulation of megalin/LRP2 by transforming growth factor beta (TGF-ß1).

Megalin/LRP2 is a receptor that plays important roles in the physiology of several organs, such as kidney, lung, intestine, and gallbladder and also in the physiology of the nervous system. Megalin expression is reduced in diseases associated with fibrosis, including diabetic nephropathy, hepatic fibrosis and cholelithiasis, as well as in some breast and prostate cancers. One of the hallmarks of these conditions is the presence of the cytokine transforming growth factor beta (TGF-ß). Although TGF-ß has been implicated in the reduction of megalin levels, the molecular mechanism underlying this regulation is not well understood. Here, we show that treatment of two epithelial cell lines (from kidney and gallbladder) with TGF-ß1 is associated with decreased megalin mRNA and protein levels, and that these effects are reversed by inhibiting the TGF-ß1 type I receptor (TGF-ßRI). Based on in silico analyses, the two SMAD-binding elements (SBEs) in the megalin promoter are located at positions -57 and -605. Site-directed mutagenesis of the SBEs and chromatin immunoprecipitation (ChIP) experiments revealed that SMAD2/3 transcription factors interact with SBEs. Both the presence of SMAD2/3 and intact SBEs were associated with repression of the megalin promoter, in the absence as well in the presence of TGF-ß1. Also, reduced megalin expression and promoter activation triggered by high concentration of albumin are dependent on the expression of SMAD2/3. Interestingly, the histone deacetylase inhibitor Trichostatin A (TSA), which induces megalin expression, reduced the effects of TGF-ß1 on megalin mRNA levels. These data show the significance of TGF-ß and the SMAD2/3 signalling pathway in the regulation of megalin and explain the decreased megalin levels observed under conditions in which TGF-ß is upregulated, including fibrosis-associated diseases and cancer.