DOI: 10.60816/gwe9-y861
title: Amyotrophic Lateral Sclerosis and swim training affect copper metabolism in skeletal muscle in a mouse model of disease

description: <p style="line-height:1.5;margin-top:1.2rem;margin-bottom:1.2rem;color:rgb( 33 , 33 , 33 );font-family:&#39;blinkmacsystemfont&#39; , &#39;-apple-system&#39; , &#39;segoe ui&#39; , &#39;roboto&#39; , &#39;oxygen&#39; , &#39;ubuntu&#39; , &#39;cantarell&#39; , &#39;fira sans&#39; , &#39;droid sans&#39; , &#39;helvetica neue&#39; , sans-serif;font-size:16px;letter-spacing:normal"><strong style="font-weight:bold">Introduction/aims: </strong>Swim training and regulation of copper metabolism result in clinical benefits in amyotrophic lateral sclerosis (ALS) mice. Therefore, the study aimed to determine whether swim training improves copper metabolism by modifying copper metabolism in the skeletal muscles of ALS mice.</p><p style="line-height:1.5;margin-top:1.2rem;margin-bottom:1.2rem;color:rgb( 33 , 33 , 33 );font-family:&#39;blinkmacsystemfont&#39; , &#39;-apple-system&#39; , &#39;segoe ui&#39; , &#39;roboto&#39; , &#39;oxygen&#39; , &#39;ubuntu&#39; , &#39;cantarell&#39; , &#39;fira sans&#39; , &#39;droid sans&#39; , &#39;helvetica neue&#39; , sans-serif;font-size:16px;letter-spacing:normal"><strong style="font-weight:bold">Methods: </strong>SOD1G93A mice (n &#61; 6 per group) were used as the ALS model, and wild-type B6SJL (WT) mice as controls (n &#61; 6). Mice with ALS were analyzed before the onset of ALS (ALS BEFORE), at baseline ALS (first disease symptoms, trained and untrained, ALS ONSET), and at the end of ALS (last stage disease, trained and untrained, ALS TERMINAL). Copper concentrations and the level of copper metabolism proteins in the skeletal muscles of the lower leg were determined.</p><p style="line-height:1.5;margin-top:1.2rem;margin-bottom:1.2rem;color:rgb( 33 , 33 , 33 );font-family:&#39;blinkmacsystemfont&#39; , &#39;-apple-system&#39; , &#39;segoe ui&#39; , &#39;roboto&#39; , &#39;oxygen&#39; , &#39;ubuntu&#39; , &#39;cantarell&#39; , &#39;fira sans&#39; , &#39;droid sans&#39; , &#39;helvetica neue&#39; , sans-serif;font-size:16px;letter-spacing:normal"><strong style="font-weight:bold">Results: </strong>ALS disease caused a reduction in the copper concentration in ALS TERMINAL untrained mice compared with the ALS BEFORE (10.43 ± 1.81 and 38.67 ± 11.50 μg/mg, respectively, p &#61; .0213). The copper chaperon for SOD1 protein, which supplies copper to SOD1, and ATPase7a protein (copper exporter), increased at the terminal stage of disease by 57% (p &#61; .0021) and 34% (p &#61; .0372), while the CTR1 protein (copper importer) decreased by 45% (p &#61; .002). Swim training moderately affected the copper concentration and the concentrations of proteins responsible for copper metabolism in skeletal muscles.</p><p style="line-height:1.5;margin-top:1.2rem;margin-bottom:1.2rem;color:rgb( 33 , 33 , 33 );font-family:&#39;blinkmacsystemfont&#39; , &#39;-apple-system&#39; , &#39;segoe ui&#39; , &#39;roboto&#39; , &#39;oxygen&#39; , &#39;ubuntu&#39; , &#39;cantarell&#39; , &#39;fira sans&#39; , &#39;droid sans&#39; , &#39;helvetica neue&#39; , sans-serif;font-size:16px;letter-spacing:normal"><strong style="font-weight:bold">Discussion: </strong>The results show disturbances in skeletal muscle copper metabolism associated with ALS progression, which is moderately affected by swim training. From a clinical point of view, exercise in water for ALS patients should be an essential element of rehabilitation for maintaining quality of life.</p>
description: <p class="MsoNormal" style="text-align:justify"><span lang="EN-GB" style="font-family:&#39;times new roman&#39; , serif">Animals</span></p><p class="MsoNormal" style="text-align:justify"><span lang="EN-GB" style="font-family:&#39;times new roman&#39; , serif">SOD1G93A
mice (n &#61; 6 per group) were used as the ALS model, and wild-type B6SJL (WT)
mice as controls (n &#61; 6). Mice with ALS were analyzed before the onset of ALS
(ALS BEFORE), at baseline ALS (first disease symptoms, trained and untrained,
ALS ONSET), and at the end of ALS (last stage disease, trained and untrained,
ALS TERMINAL). The animals in the ALS BEFORE group were euthanized at the
10th week of age. The ALS ONSET mice were euthanized when the first disease
symptoms were observed in untrained animals (16th week of age). The ALS
TERMINAL groups were euthanized when the last stage of disease was observed in
untrained animals (18th to 19th week of life).</span></p><p class="MsoNormal" style="text-align:justify"><span lang="EN-GB" style="font-family:&#39;times new roman&#39; , serif">Methods</span></p><p class="MsoNormal" style="text-align:justify"><span lang="EN-GB" style="font-family:&#39;times new roman&#39; , serif">Copper
levels in skeletal muscle homogenates were analyzed with ICP-OES (Ciros Vision,
SPECTRO Analytical Instruments Kleve, Germany) after wet-washing samples with
65% nitric acid as previously described. The copper concentration in the
skeletal muscle homogenates was expressed as micrograms per milligram of
protein.</span></p><p class="MsoNormal" style="text-align:justify"><span lang="EN-GB" style="font-family:&#39;times new roman&#39; , serif">The level
of copper metabolism proteins in the skeletal muscles of the lower leg were assessed
by WB method. Only Metallothionein 1 protein level was determined with
enzyme-linked immunosorbent assay (ELISA) according to the method described in
the Datasheet for ELK2455 Mouse MT1 (Metallothionein 1) ELISA Kit (ELK
Biotechnology, Wuhan, China).</span></p><p class="MsoNormal" style="text-align:justify"><span lang="EN-GB" style="font-family:&#39;times new roman&#39; , serif">Proteomic
analysis. The levels of 15 investigated proteins (membrane primary amine
oxidase, AOC3; metalloreductase, STEAP3; glutaredoxin-like protein, C5orf63
homolog; ceruloplasmin, copper transport protein, ATOX1; superoxide dismutase
1, SOD1; cytochrome c oxidase assembly protein, SCO1; cytochrome c oxidase
subunit 1; cytochrome c oxidase subunit 2; copper homeostasis protein cut C
homolog; lysyl oxidase homolog 1, LOXL1; cytochrome c oxidase assembly protein,
COX11; cytochrome c oxidase copper chaperone, COX17; copper chaperone for
superoxide dismutase, CCS; and major prion protein, PRIO) were determined in
the BEFORE ALS and TERMINAL ALS untrained and trained groups using liquid
chromatography MS3 spectrometry (LC–MS/MS) at the Thermo Fisher Center for
Multiplexed Proteomics (Department of Cell Biology, Harvard Medical School,
Cambridge, MA, USA).</span></p>