Choline administration attenuates aspects of the dystrophic pathology in mdx mice Francesca M. Alves a, b , Marissa K. Caldow a, b , Jennifer Trieu a, b , Tim Naim a, b , Magdalene K. Montgomery b , Matthew J. Watt b , Gordon S. Lynch a, b, 1 , Ren  e Koopman a, b, *, 1 a Centre for Muscle Research, The University of Melbourne, Australia b Department of Physiology, The University of Melbourne, Australia article info Article history: Received 3 December 2018 Accepted 22 December 2018 Available online 29 December 2018 Keywords: Choline Dystrophy mdx mice Diaphragm Fibrosis summary Background & aims: Dystrophic muscle fibres are fragile and prone to breakage, leading to impaired Ca 2þ homeostasis and excessive inflammation, resulting in muscle wasting and weakness. Choline, an essential water-soluble nutrient, is involved in multiple biolog- ical processes, including modulation of inflammation and oxidative stress. We tested the hypothesis that choline supplementation would ameliorate the dystrophic pathology in mdx mice. Methods: Three-week old male mdx mice (n ¼ 40) were fed control purified laboratory chow (CON; n ¼ 20) or a choline-enriched diet (5 g/kg choline; CHL, n ¼ 20) for four weeks. Rotarod performance, grip strength and running (wheel) distance were assessed during treatment. Markers of Ca 2þ -handling, inflammation, oxidative stress and fibrosis were measured in the diaphragm, quadriceps muscle and the liver. Results: Choline-treated mdx mice displayed less macrophage (CD68 -33%, P < 0.05) and collagen infiltration (34%, P < 0.05) and reduced Tgfb3 mRNA expression (P < 0.05) in the diaphragm compared to untreated mdx mice. Choline supplementation increased maximal SERCA activity (38%, P < 0.05) and reduced markers of inflammatory (Tnfa, F4/80 and Cd206 mRNA, P < 0.05) processes compared with untreated mdx mice. In the liver there Abbreviations: DMD, Duchenne muscular dystrophy; TMAO, Trimethylamine oxide; AST, Aspartate aminotransferase; ALT, Alanine aminotransferase. * Corresponding author. Centre for Muscle Research, The University of Melbourne, Melbourne, VIC 3010 Australia. Fax: þ61 3 8344 5818. E-mail address: rkoopman@unimelb.edu.au (R. Koopman). 1 These authors contributed equally. Contents lists available at ScienceDirect Clinical Nutrition Experimental journal homepage: http:// www.clinicalnutritionexperimental.com https://doi.org/10.1016/j.yclnex.2018.12.005 2352-9393/© 2018 The Authors. Published by Elsevier Ltd on behalf of European Society for Clinical Nutrition and Metabolism. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Clinical Nutrition Experimental 24 (2019) 83e91