Jeremie Zander Lindeque1, Juan Hidalgo2, Roan Louw1, Francois Hendrikus? Van Der Westhuizen1

SYSTEMIC AND ORGAN SPECIFIC METABOLIC VARIATION IN METALLOTHIONEIN KNOCKOUT MICE CHALLENGED WITH SWIMMING EXERCISE.

Jeremie Zander Lindeque1, Juan Hidalgo2, Roan Louw1, Francois Hendrikus? van der Westhuizen1

1Centre for Human Metabonomics, North-West University (Potchefstroom Campus), South Africa.

2Institute of Neurosciences, Department of Cellular Biology, Physiology and Immunology, Faculty of Biosciences, Autonomous University of Barcelona, Spain.

Index

Sample preparation
Metabolic trends as determined with ASCA.
Tables with metabolic differences
Additional Figures

Sample preparation

Fig. S-1: Sample preparation and analytical procedures for liver (A), plasma (B), brain and gastrocnemius (C) samples. A single biphasic metabolite extraction was used on all tissue samples. All tissue extracts were prepared for LC-MS analysis and GC-MS analysis of trimethylsilyl esters. The liver extract was also prepared for the GC-MS analysis of FAMEs. Deproteinized plasma was used for LC-MS analysis only. ACN, acetonitrile.

Metabolic trends as determined with ASCA.

Fig. S-2:Most prominent patterns observed in the data as determined by ASCA.The variance pattern of the variables contributing to component 1 which did not show interaction is shown in the top figures for all the studied tissues (A). The variance pattern of the variables that showed interaction between the groups is shown in the bottom plots (B).

Tables with metabolic differences

Table S-1a: Metabolite levels significantly different between the MT1+2KO and WT mice during unchallenged conditions

Class / Metabolite / ID # / Plasma / Gastrocnemius / Liver / Brain
Direction# / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p)
Amino acid metabolism / Valine / HMDB00883 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.989 / 0.033
Alanine / HMDB00161 / N.D. / N.S. / N.D. / N.S. / – / 0.879 / 0.047 / + / 1.326 / 0.012
Aspartic Acid / HMDB00191 / N.D. / N.S. / N.D. / N.S. / + / 1.247 / 0.013 / N.D. / N.S.
Citrulline / HMDB00904 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.090 / 0.022
Lysine / HMDB00182 / + / 1.078 / 0.019 / + / 1.526 / 0.002 / N.D. / N.S. / N.D. / N.S.
Phenylpyruvic acid / HMDB00205 / N.D. / N.S. / + / 1.328 / 0.025 / N.D. / N.S. / N.D. / N.S.
Tyrosine / HMDB00158 / N.D. / N.S. / + / 1.422 / 0.033 / N.D. / N.S. / N.D. / N.S.
2-Aminoacrylic acid / HMDB03609 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.498 / 0.012
Sarcosine / HMDB00271 / N.D. / N.S. / N.D. / N.S. / + / 1.017 / 0.020 / N.D. / N.S.
Antioxidant metabolism / Pyroglutamic acid / HMDB00267 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.892 / 0.001
Carbohydrate metabolism / ? Fructose / HMDB00660 / N.D. / N.S. / N.D. / N.S. / + / 0.880 / 0.029 / N.D. / N.S.
? Glucose / HMDB00122 / N.D. / N.S. / N.D. / N.S. / + / 1.113 / 0.015 / N.D. / N.S.
? Galactose / HMDB00143 / N.D. / N.S. / N.D. / N.S. / + / 0.898 / 0.041 / N.D. / N.S.
? Fucose / HMDB00174 / N.D. / N.S. / N.D. / N.S. / + / 0.916 / 0.010 / N.D. / N.S.
? Sucrose / HMDB00258 / N.D. / N.S. / N.D. / N.S. / + / 1.104 / 0.004 / N.D. / N.S.
? Sorbitol / HMDB00247 / N.D. / N.S. / N.D. / N.S. / + / 1.116 / 0.007 / N.D. / N.S.
Mannose-6-phosphate / HMDB01078 / N.D. / N.S. / N.D. / N.S. / + / 1.430 / 0.002 / N.D. / N.S.
Lipid metabolism / Glycerol 3-phosphate / HMDB00126 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.918 / 0.038
Bound fatty acids / Pentadecanoic acid / HMDB00826 / N.D. / N.S. / N.D. / N.S. / + / 1.056 / 0.004 / N.D. / N.S.
10E-heptadecenoic acid / HMDB02259 / N.D. / N.S. / N.D. / N.S. / + / 1.190 / 0.011 / N.D. / N.S.
Oleic acid / HMDB00207 / N.D. / N.S. / N.D. / N.S. / + / 0.909 / 0.022 / N.D. / N.S.
Eicosenoic acid / HMDB02231 / N.D. / N.S. / N.D. / N.S. / + / 1.330 / 0.010 / N.D. / N.S.
Free fatty acids / Eicosanoic acid / HMDB02212 / N.D. / N.S. / – / 0.990 / 0.010 / N.D. / N.S. / N.D. / N.S.
Heneicosanoic acid / HMDB02345 / N.D. / N.S. / – / 0.963 / 0.017 / N.D. / N.S. / N.D. / N.S.
Margaric acid / HMDB02259 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.373 / 0.009
Palmitoleic acid / HMDB03229 / N.D. / N.S. / N.D. / N.S. / – / 1.240 / 0.006 / N.D. / N.S.
Hydroxyadipic acid / HMDB00321 / N.D. / N.S. / N.D. / N.S. / + / 0.820 / 0.028 / N.D. / N.S.
Myristic acid / HMDB00806 / – / 0.930 / 0.035 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Phospholipid metabolism / CDPcholine / HMDB01413 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.834 / 0.004
Ethanolamine / HMDB00149 / N.D. / N.S. / N.D. / N.S. / + / 1.702 / 0.002 / N.D. / N.S.
Phosphocholine / HMDB01565 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.934 / 0.003
Other lipids / 1-Monopalmitin / HMDB11564 / N.D. / N.S. / – / 1.607 / 0.003 / N.D. / N.S. / N.D. / N.S.
Neurotransmitters / N-acetylaspartate / HMDB00812 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.736 / <0.001
Pyrimidine metabolism / Cytidine / HMDB00089 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 2.364 / <0.001
Uracil / HMDB00300 / N.D. / N.S. / N.D. / N.S. / + / 1.521 / <0.001 / + / 1.112 / 0.020
Uridine / HMDB00296 / N.D. / N.S. / N.D. / N.S. / + / 0.971 / 0.013 / + / 1.309 / 0.022
TCA metabolism / Fumaric acid / HMDB00134 / N.D. / N.S. / N.D. / N.S. / + / 1.414 / 0.004 / N.D. / N.S.
Lactic acid / HMDB00190 / N.D. / N.S. / + / 1.019 / 0.027 / N.D. / N.S. / N.D. / N.S.
Malic acid / HMDB00156 / N.D. / N.S. / N.D. / N.S. / + / 1.090 / 0.007 / N.D. / N.S.
Crotonic acid / HMDB10720 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.797 / 0.004
Vitamin metabolism / Corrinoid / HMDB04269 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.264 / 0.010
α-tocopheronolactone / HMDB01893 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.819 / 0.004
Other / Gluconic acid / HMDB00625 / N.D. / N.S. / N.D. / N.S. / + / 1.087 / 0.049 / N.D. / N.S.
Oxalic acid / HMDB02329 / N.D. / N.S. / N.D. / N.S. / – / 1.328 / 0.034 / N.D. / N.S.
Aminomalonic acid / HMDB01147 / N.D. / N.S. / N.D. / N.S. / – / 0.934 / 0.039 / N.D. / N.S.
# The direction indicates whether a compound was higher (+) or lower (-) in the MT knockout mice in comparison to the WT mice
A bold p-value signifies that a significant difference was seen between the MT1+2KO and MT3KO mice.
N.D., Not determined / N.S., Not significant

Table S-1b: Metabolite levels significantly different between the MT3KO and WT mice during unchallenged conditions

Table 2a: Metabolite levels significantly different between the MT1+2KO and WT after challenged with exercise

Class / Metabolite / ID # / Plasma / Gastrocnemius / Liver / Brain
Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p)
Amino acid metabolism / Glutamate / HMDB00148 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.327 / 0.013
2-Ketobutyric acid / HMDB00005 / N.D. / N.S. / N.D. / N.S. / + / 1.005 / 0.045 / N.D. / N.S.
2-Keto-6-acetamidocaproate / HMDB12150 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.212 / 0.007
2-Pyrrolidone-5-carboxylic acid / HMDB00805 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.608 / < 0.001
Alanine / HMDB00161 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.394 / < 0.001
Aspartic acid / HMDB00191 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.273 / < 0.001
Cadaverine / HMDB02322 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.908 / < 0.001
Glutamine / HMDB00641 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.217 / < 0.001
Isoleucine / HMDB00172 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.802 / < 0.001
Phenylethylamine / HMDB12275 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.787 / < 0.001
Proline / HMDB00162 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.148 / 0.004
Serine / HMDB00187 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.193 / < 0.001
Threonine / HMDB00167 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.387 / < 0.001
Valine / HMDB00883 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.865 / < 0.001
Pyrroline-3-hydroxy-5-carboxylate / HMDB01369 / – / 1.323 / 0.020 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Antioxidant metabolism / Pyroglutamic acid / HMDB00267 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 2.382 / < 0.001
Carbohydrate metabolism / ? Fructose / HMDB00660 / N.D. / N.S. / – / 1.026 / 0.004 / N.D. / N.S. / + / 1.587 / < 0.001
? Ribose / HMDB00283 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.181 / < 0.001
? Erythrose / HMDB02649 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.174 / < 0.001
? Sedoheptulose / HMDB03219 / N.D. / N.S. / N.D. / N.S. / + / 0.936 / 0.019 / N.D. / N.S.
Creatine phosphate system / Creatinine / HMDB00562 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.828 / 0.001
Hormone metabolism / Corticosterone / HMDB01547 / + / 0.940 / 0.040 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Lipid metabolism / Carnitine / HMDB00562 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.026 / 0.045
Free fatty acids / Eicosanoic acid / HMDB02212 / N.D. / N.S. / – / 2.398 / 0.001 / N.D. / N.S. / + / 2.623 / < 0.001
Glycerol / HMDB00131 / N.D. / N.S. / N.D. / N.S. / + / 0.861 / 0.009 / + / 1.530 / < 0.001
Heneicosanoic acid / HMDB02345 / N.D. / N.S. / – / 1.908 / 0.002 / N.D. / N.S. / + / 1.177 / < 0.001
Margaric acid / HMDB02259 / N.D. / N.S. / – / 1.343 / 0.025 / N.D. / N.S. / + / 2.829 / < 0.001
Stearic acid / HMDB00827 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.814 / 0.001
Palmitoleic acid / HMDB03229 / + / 1.292 / 0.003 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Hexadecenoyl carnitine / HMDB00222 / + / 1.099 / 0.009 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Bound fatty acids / Eicosenoic acid / HMDB02231 / N.D. / N.S. / N.D. / N.S. / + / 0.802 / 0.008 / N.D. / N.S.
Acylglycerides / Monopalmitin / HMDB11564 / N.D. / N.S. / – / 1.313 / 0.037 / N.D. / N.S. / + / 0.869 / 0.007
Monostearin / HMDB11131 / N.D. / N.S. / – / 1.628 / 0.011 / N.D. / N.S. / + / 0.914 / 0.010
Phospholipid metabolism / Ethanolamine / HMDB00149 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.469 / < 0.001
Glycerophosphocholine / HMDB00086 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 0.921 / 0.018
Myo-inositol / HMDB00211 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.852 / < 0.001
Phosphocholine / HMDB01565 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.327 / 0.006
PE(18:2/18:2) / HMDB09093 / – / 2.018 / 0.001 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Other lipids / Cholesterol / HMDB00067 / N.D. / N.S. / N.D. / N.S. / + / 1.515 / 0.003 / N.D. / N.S.
Glyceric acid / HMDB06372 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.935 / 0.001
Neurotransmitters / N-acetylaspartate / HMDB00812 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 2.701 / < 0.001
γ-Aminobutyric acid / HMDB00112 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.945 / 0.001
Purine metabolism / Adenine / HMDB00034 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.241 / < 0.001
AMP / HMDB00045 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.197 / 0.001
Ribitol / HMDB00508 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.679 / < 0.001
Pyrimidine metabolism / Uracil / HMDB00300 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.449 / 0.007
TCA metabolism / 2-Ketoglutaric acid / HMDB00208 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.214 / 0.007
3-Hydroxybutyric acid / HMDB00357 / N.D. / N.S. / N.D. / N.S. / + / 0.893 / 0.002 / N.D. / N.S.
Fumaric acid / HMDB00134 / N.D. / N.S. / N.D. / N.S. / + / 0.991 / 0.032 / + / 1.820 / < 0.001
Lactic acid / HMDB00190 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.916 / < 0.001
Malic acid / HMDB00156 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.564 / < 0.001
Succinic acid / HMDB00254 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.862 / 0.018
Other / Gluconic acid / HMDB00625 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 0.985 / < 0.001
Aminomalonic acid / HMDB01147 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.353 / 0.001
Oxalic acid / HMDB02329 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.325 / < 0.001

Table 2b: Metabolite levels significantly different between the MT3KO and WT after challenged with exercise

Class / Metabolite / ID # / Plasma / Gastrocnemius / Liver / Brain
Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p) / Direction / Effect size (d) / ANOVA (p)
Amino acid metabolism / Alanine / HMDB00161 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.644 / < 0.001
Aspartic acid / HMDB00191 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.326 / < 0.001
Cadaverine / HMDB02322 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.090 / < 0.001
Glutamine / HMDB00641 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.193 / < 0.001
Isoleucine / HMDB00172 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.134 / < 0.001
Phenylalanine / HMDB00159 / N.D. / N.S. / + / 1.891 / 0.033 / N.D. / N.S. / N.D. / N.S.
Pyrroline-3-hydroxy-5-carboxylate / HMDB01369 / – / 0.946 / 0.020 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Threonine / HMDB00167 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.163 / < 0.001
Valine / HMDB00883 / N.D. / N.S. / N.D. / N.S. / + / 1.054 / 0.007 / – / 0.949 / < 0.001
2-Methylbutyrylcarnitine / HMDB00378 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / + / 1.185 / 0.042
Antioxidant metabolism / Pyroglutamic acid / HMDB00267 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.321 / < 0.001
Carbohydrate metabolism / ? Fructose / HMDB00660 / N.D. / N.S. / + / 1.407 / 0.004 / N.D. / N.S. / – / 1.601 / < 0.001
? Ribose / HMDB00283 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.160 / < 0.001
? Sedoheptulose / HMDB03219 / N.D. / N.S. / N.D. / N.S. / + / 1.217 / 0.019 / N.D. / N.S.
? Erythrose / HMDB02649 / N.D. / N.S. / – / 1.835 / 0.016 / N.D. / N.S. / – / 1.207 / < 0.001
Creatine phosphate system / Creatine / HMDB00562 / N.D. / N.S. / N.D. / N.S. / + / 0.923 / 0.009 / N.D. / N.S.
Creatinine / HMDB00562 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.009 / 0.001
Lipid metabolism / Arachidonic acid / HMDB01043 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.407 / 0.006
Free fatty acids / Eicosanoic acid / HMDB02212 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 2.439 / < 0.001
Heneicosanoic acid / HMDB02345 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.241 / < 0.001
Margaric acid / HMDB02259 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.767 / < 0.001
Stearic acid / HMDB00827 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 0.953 / 0.001
Bound fatty acids / Eicosenoic acid / HMDB02231 / N.D. / N.S. / N.D. / N.S. / + / 0.963 / 0.008 / N.D. / N.S.
γ-Linolenic acid / HMDB03073 / N.D. / N.S. / N.D. / N.S. / + / 0.885 / 0.047 / N.D. / N.S.
Phospholipid metabolism / Myo-inositol / HMDB00211 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 2.649 / < 0.001
PC(16:0/16:0) / HMDB00564 / + / 0.854 / 0.039 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
PE(18:2/18:2) / HMDB09093 / – / 1.212 / 0.001 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S.
Phosphocholine / HMDB01565 / N.D. / N.S. / N.D. / N.S. / + / 0.916 / 0.014 / – / 1.650 / 0.006
Ethanolamine / HMDB00149 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 0.803 / < 0.001
Glycerophosphocholine / HMDB00086 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.181 / 0.018
Other lipids / Cholesterol / HMDB00067 / N.D. / N.S. / N.D. / N.S. / + / 1.137 / 0.003 / N.D. / N.S.
Glycerol / HMDB00131 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.416 / < 0.001
Neurotransmitters / N-acetylaspartate / HMDB00812 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.995 / < 0.001
γ-Aminobutyric acid / HMDB00112 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.012 / 0.001
Purine metabolism / Inosine / HMDB00195 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 0.937 / 0.003
Pyrimidine metabolism / Uracil / HMDB00300 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.215 / 0.007
Uridine / HMDB00296 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.025 / 0.008
TCA metabolism / Acetic acid / HMDB00042 / N.D. / N.S. / N.D. / N.S. / + / 1.290 / 0.020 / N.D. / N.S.
Fumaric acid / HMDB00134 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 0.956 / < 0.001
Malic acid / HMDB00156 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 0.887 / < 0.001
Other / Oxalic acid / HMDB02329 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.348 / < 0.001
Gluconic acid / HMDB00625 / N.D. / N.S. / N.D. / N.S. / N.D. / N.S. / – / 1.174 / < 0.001

Table S-3: Circulating metabolite levels significantly different in all three strains of mice that swam when compared to their respective controls.

Group / Metabolite / ID # / WT / MT1+2KO / MT3KO
Direction# / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p)
Amino acid metabolism / Alanine / HMDB00161 / + / 0.974 / 0.038 / N.D. / N.S. / + / 1.404 / 0.024
Leucine / HMDB00687 / – / 1.189 / 0.009 / – / 1.592 / < 0.001 / N.D. / N.S.
Methylglutaconic acid / HMDB00522 / N.D. / N.S. / – / 1.086 / 0.004 / N.D. / N.S.
Methionine / HMDB00696 / – / 0.855 / 0.034 / – / 1.394 / < 0.001 / N.D. / N.S.
N-Acetylglutamine / HMDB06029 / + / 0.827 / 0.040 / N.D. / N.S. / N.D. / N.S.
Phenylalanine / HMDB00159 / – / 1.240 / 0.003 / – / 1.260 / 0.002 / N.D. / N.S.
Phenylpyruvic acid / HMDB00205 / – / 1.302 / 0.001 / – / 1.386 / < 0.001 / N.D. / N.S.
Proline / HMDB00162 / – / 1.133 / 0.008 / – / 1.222 / 0.001 / N.D. / N.S.
Threonine / HMDB00167 / N.D. / N.S. / – / 1.324 / 0.001 / N.D. / N.S.
Tryptophan / HMDB00929 / N.D. / N.S. / – / 1.413 / < 0.001 / N.D. / N.S.
Xanthurenic acid / HMDB00881 / + / 1.142 / 0.008 / N.D. / N.S. / N.D. / N.S.
Tyrosine / HMDB00158 / – / 1.229 / 0.002 / – / 1.389 / < 0.001 / N.D. / N.S.
Valine / HMDB00883 / – / 0.856 / 0.038 / – / 1.099 / 0.009 / N.D. / N.S.
Propionyl-CoA / HMDB01275 / + / 1.015 / 0.019 / N.D. / N.S. / N.D. / N.S.
Carbohydrates / Glucose / HMDB00122 / N.D. / N.S. / – / 1.175 / 0.005 / N.D. / N.S.
Cofactor metabolism / NADPH / HMDB00221 / + / 1.155 / 0.014 / N.D. / N.S. / N.D. / N.S.
Niacinamide / HMDB01406 / + / 0.840 / 0.024 / N.D. / N.S. / N.D. / N.S.
Creatine phosphate system / Creatinine / HMDB00562 / + / 1.089 / 0.022 / N.D. / N.S. / N.D. / N.S.
Hormones / Corticosterone / HMDB01547 / + / 1.286 / 0.005 / + / 3.419 / < 0.001 / N.D. / N.S.
Lipid metabolism / Carnitine / HMDB00562 / N.D. / N.S. / – / 1.143 / 0.003 / N.D. / N.S.
Free fatty acids / 3-Oxohexadecanoic acid / HMDB10733 / + / 0.947 / 0.033 / N.D. / N.S. / N.D. / N.S.
9,10-Dihydroxy-12-octadecenoic acid / HMDB04704 / N.D. / N.S. / N.D. / N.S. / – / 1.273 / 0.040
Linoleic Acid / HMDB05047 / N.D. / N.S. / + / 1.937 / < 0.001 / N.D. / N.S.
Palmitoleic acid / HMDB03229 / + / 1.273 / 0.003 / + / 0.990 / 0.012 / + / 1.259 / 0.032
Tetradecenoic acid / HMDB00499 / N.D. / N.S. / N.D. / N.S. / + / 1.503 / 0.037
2,3-Dihydroxy stearic acid / N.D. / N.S. / N.D. / N.S. / – / 1.303 / 0.045
Bile & Cholesterols / Chenodeoxycholic acid / HMDB00518 / N.D. / N.S. / N.D. / N.S. / + / 4.181 / < 0.001
Chenodeoxycholic acid 3-glucuronide / HMDB00518 / + / 1.596 / 0.001 / + / 1.437 / 0.003 / N.D. / N.S.
Taurocholic acid / HMDB00922 / + / 1.544 / 0.002 / N.D. / N.S. / N.D. / N.S.
Acylglycerides / DG(16:0/18:1/0:0) / HMDB07101 / + / 0.923 / 0.026 / N.D. / N.S. / N.D. / N.S.
DG(18:3/18:3/0:0) / LMGL02010079 / N.D. / N.S. / + / 1.072 / 0.007 / N.D. / N.S.
Phospholipids / LysoPC(16:0) / HMDB10382 / – / 0.948 / 0.045 / N.D. / N.S. / N.D. / N.S.
LysoPC(16:1) / HMDB10383 / N.D. / N.S. / N.D. / N.S. / – / 1.496 / 0.025
LysoPC(18:2) / HMDB10386 / – / 1.054 / 0.030 / N.D. / N.S. / N.D. / N.S.
LysoPC(18:4) / HMDB10389 / N.D. / N.S. / + / 0.889 / 0.032 / N.D. / N.S.
LysoPC(22:5) / HMDB10403 / – / 1.440 / 0.005 / N.D. / N.S. / N.D. / N.S.
LysoPE(0:0/18:2) / HMDB11477 / N.D. / N.S. / – / 1.198 / 0.005 / N.D. / N.S.
LysoPE(16:0/0:0) / HMDB11503 / N.D. / N.S. / – / 1.487 / 0.001 / – / 2.201 / 0.002
PA(10:0/10:0) / LMGP10010022 / + / 1.500 / 0.001 / N.D. / N.S. / N.D. / N.S.
PA(10:0/10:0) / LMGP10010022 / N.D. / N.S. / + / 1.330 / 0.003 / N.D. / N.S.
PC(18:0/24:1) / HMDB08059 / N.D. / N.S. / + / 1.333 / 0.002 / N.D. / N.S.
PC(18:0/P-18:0) / N.D. / N.S. / N.D. / N.S. / + / 1.196 / 0.050
PC(20:0/P-18:0) / N.D. / N.S. / N.D. / N.S. / + / 1.921 / 0.007
PC(20:0/P-18:1) / HMDB08293 / N.D. / N.S. / N.D. / N.S. / + / 1.265 / 0.040
PC(22:0/P-18:1) / HMDB08557 / N.D. / N.S. / N.D. / N.S. / + / 1.879 / 0.011
PC(O-1:0/2:0) / LMGP01020005 / + / 1.598 / 0.001 / N.D. / N.S. / N.D. / N.S.
PC(O-16:1/2:0) / LMGP01020147 / – / 1.103 / 0.016 / N.D. / N.S. / N.D. / N.S.
PE(18:2/18:2) / HMDB09093 / N.D. / N.S. / – / 1.137 / 0.008 / N.D. / N.S.
PE(19:0/22:0) / N.D. / N.S. / N.D. / N.S. / + / 1.242 / 0.041
PE(20:0/23:0) / LMGP02010076 / N.D. / N.S. / N.D. / N.S. / + / 1.460 / 0.027
PG(10:0/10:0) / LMGP04010032 / + / 1.070 / 0.021 / + / 1.056 / 0.021 / N.D. / N.S.
PG(16:0/0:0) / LMGP04050008 / N.D. / N.S. / + / 1.310 / 0.005 / N.D. / N.S.
PG(18:1/0:0) / LMGP04050006 / + / 1.938 / < 0.001 / N.D. / N.S. / N.D. / N.S.
Other lipids / N-Oleoyl-L-Serine / N.D. / N.S. / + / 0.950 / 0.022 / N.D. / N.S.
MGDG(18:5/18:5) / LMGL05010012 / N.D. / N.S. / + / 1.196 / 0.011 / N.D. / N.S.
Purine metabolism / AMP / HMDB00045 / + / 0.919 / 0.039 / N.D. / N.S. / N.D. / N.S.
Hypoxanthine / HMDB00157 / + / 1.340 / 0.005 / + / 0.883 / 0.024 / + / 1.801 / 0.008
Pyrimidine metabolism / dUMP / HMDB01409 / N.D. / N.S. / – / 0.863 / 0.032 / N.D. / N.S.
Uridine / HMDB00296 / N.D. / N.S. / + / 0.960 / 0.025 / N.D. / N.S.
TCA metabolism / Lactic acid / HMDB00190 / N.D. / N.S. / + / 0.936 / 0.010 / N.D. / N.S.
Acetylcarnitine / HMDB00201 / + / 1.409 / 0.001 / N.D. / N.S. / N.D. / N.S.
Other / 3-Deoxyvitamin D3 / N.D. / N.S. / – / 0.889 / 0.032 / N.D. / N.S.
Glucaric acid lactone / HMDB00663 / N.D. / N.S. / N.D. / N.S. / + / 1.704 / 0.015
# The direction indicates whether a compound was higher (+) or lower (-) in the exercise group in comparison to the control. N.D., Not determined; N.S., Not significant

Table S-4: Metabolite levels different in the gastrocnemius of all three strains of mice that swam when compared to their respective controls

Group / Metabolite / ID # / WT / MT1+2KO / MT3KO
Direction / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p)
Amino acid metabolism / Ketoisovaleric acid / HMDB00019 / N.D. / N.S. / – / 0.915 / 0.051 / N.D. / N.S.
3-Hydroxyisovalerylcarnitine / HMDB06870 / N.D. / N.S. / – / 0.921 / 0.040 / N.D. / N.S.
Lysine / HMDB00182 / N.D. / N.S. / – / 1.886 / 0.001 / N.D. / N.S.
Methionine / HMDB00696 / N.D. / N.S. / – / 2.057 / < 0.001 / N.D. / N.S.
Phenylalanine / HMDB00159 / N.D. / N.S. / – / 1.753 / 0.001 / N.D. / N.S.
Phenylpyruvic acid / HMDB00205 / N.D. / N.S. / – / 2.185 / < 0.001 / N.D. / N.S.
Proline / HMDB00162 / N.D. / N.S. / N.D. / N.S. / + / 1.488 / 0.038
Tyrosine / HMDB00158 / N.D. / N.S. / – / 2.207 / < 0.001 / N.D. / N.S.
5'-Methylthioadenosine / HMDB01173 / N.D. / N.S. / – / 0.930 / 0.033 / N.D. / N.S.
Antioxidant metabolism / Cysteinylglycine / HMDB00078 / N.D. / N.S. / – / 0.956 / 0.029 / N.D. / N.S.
Glutathione / HMDB00125 / N.D. / N.S. / – / 1.288 / 0.011 / N.D. / N.S.
Carbohydrates / Erythrose / HMDB02649 / N.D. / N.S. / – / 0.868 / 0.018 / N.D. / N.S.
Glucose / HMDB00122 / N.D. / N.S. / – / 1.053 / 0.002 / N.D. / N.S.
Cofactor metabolism / Niacinamide / HMDB01406 / N.D. / N.S. / – / 1.733 / 0.001 / N.D. / N.S.
Creatine phosphate system / Creatine / HMDB00562 / N.D. / N.S. / – / 1.383 / 0.007 / N.D. / N.S.
Lipid metabolism / Carnitine / HMDB00562 / N.D. / N.S. / – / 2.199 / < 0.001 / N.D. / N.S.
Free fatty acids / 3-Hydroxydodecanoic acid / HMDB00387 / + / 1.028 / 0.009 / N.D. / N.S. / N.D. / N.S.
Eicosanoic acid / HMDB02212 / + / 1.961 / 0.003 / N.D. / N.S. / – / 3.272 / 0.002
Heneicosanoic acid / HMDB02345 / + / 1.471 / 0.016 / N.D. / N.S. / – / 1.679 / 0.046
Margaric acid / HMDB02259 / N.D. / N.S. / N.D. / N.S. / – / 1.693 / 0.043
Dodecanoylcarnitine / HMDB02250 / N.D. / N.S. / + / 1.162 / 0.012 / N.D. / N.S.
Elaidic carnitine / HMDB06464 / N.D. / N.S. / + / 1.448 / 0.003 / N.D. / N.S.
Hexanoylcarnitine / HMDB00756 / + / 1.284 / 0.006 / N.D. / N.S. / N.D. / N.S.
Octanoylcarnitine / HMDB00791 / + / 0.965 / 0.037 / N.D. / N.S. / N.D. / N.S.
Stearoylcarnitine / HMDB00848 / N.D. / N.S. / N.D. / N.S. / + / 1.374 / 0.037
Acylglycerides / MG(18:0/0:0/0:0) / HMDB11131 / N.D. / N.S. / N.D. / N.S. / + / 2.843 / 0.012
Phospholipid metabolism / Myo-inositol / HMDB00211 / + / 1.163 / 0.041 / N.D. / N.S. / N.D. / N.S.
PI(16:0/18:2) / HMDB09784 / N.D. / N.S. / N.D. / N.S. / + / 1.749 / 0.021
Purine metabolism / Inosine / HMDB00195 / + / 1.135 / 0.011 / N.D. / N.S. / N.D. / N.S.
Pyrimidine metabolism / Orotic acid / HMDB00226 / N.D. / N.S. / – / 1.101 / 0.017 / N.D. / N.S.
UMP / HMDB00288 / N.D. / N.S. / – / 1.065 / 0.030 / N.D. / N.S.
TCA metabolism / Acetoacetyl-CoA / HMDB01484 / N.D. / N.S. / – / 1.110 / 0.014 / N.D. / N.S.
Lactic acid / HMDB00190 / N.D. / N.S. / – / 1.723 / < 0.001 / N.D. / N.S.
Malic acid / HMDB00156 / N.D. / N.S. / N.D. / N.S. / + / 3.274 / 0.004
Acetylcarnitine / HMDB00201 / N.D. / N.S. / – / 1.566 / 0.001 / N.D. / N.S.
Other / Turanose / HMDB11740 / N.D. / N.S. / – / 1.596 / 0.004 / N.D. / N.S.

Table S-5: Metabolite levels different in the liver of all three strains of mice that swam when compared to their respective controls

Group / Metabolite / ID # / WT / MT1+2KO / MT3KO
Direction / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p)
Amino acid metabolism / 2-Ketobutyric acid / HMDB00005 / N.D. / N.S. / + / 1.433 / 0.001 / N.D. / N.S.
Alanine / HMDB00161 / N.D. / N.S. / + / 1.362 / 0.002 / N.D. / N.S.
Aspartic acid / HMDB00191 / N.D. / N.S. / N.D. / N.S. / + / 1.721 / 0.027
Cadaverine / HMDB02322 / N.D. / N.S. / + / 1.323 / 0.004 / N.D. / N.S.
Glutaric acid / HMDB00663 / N.D. / N.S. / + / 1.322 / < 0.001 / N.D. / N.S.
Glycine / HMDB00123 / N.D. / N.S. / + / 1.338 / 0.001 / N.D. / N.S.
Proline / HMDB00162 / – / 1.158 / 0.008 / N.D. / N.S. / + / 2.302 / 0.009
3-Methylglutarylcarnitine / HMDB00552 / N.D. / N.S. / + / 0.850 / 0.032 / N.D. / N.S.
Carbohydrate metabolism / Erythritol / HMDB02994 / N.D. / N.S. / N.D. / N.S. / + / 2.231 / 0.004
Glucose-6-phosphate / HMDB01401 / N.D. / N.S. / + / 1.337 / 0.002 / N.D. / N.S.
Gulonic acid / HMDB03290 / N.D. / N.S. / + / 0.862 / 0.012 / N.D. / N.S.
Creatine phosphate system / Creatine / HMDB00562 / N.D. / N.S. / – / 1.131 / 0.014 / N.D. / N.S.
Lipid metabolism / Arachidonic acid / HMDB01043 / N.D. / N.S. / N.D. / N.S. / – / 1.025 / 0.040
Bound fatty acids / Nonadecanoic acid / HMDB00772 / N.D. / N.S. / – / 1.049 / 0.017 / N.D. / N.S.
Oleic acid / HMDB00207 / N.D. / N.S. / + / 0.822 / 0.044 / N.D. / N.S.
γ-Linolenic acid / HMDB03073 / – / 0.976 / 0.003 / N.D. / N.S. / N.D. / N.S.
Free fatty acids / 13Z-Octadecenoic acid / LMFA01030290 / N.D. / N.S. / + / 1.175 / 0.002 / N.D. / N.S.
Linoleic acid / HMDB00673 / N.D. / N.S. / + / 1.040 / 0.003 / N.D. / N.S.
Nonadecanoic acid / HMDB00772 / N.D. / N.S. / N.D. / N.S. / – / 1.411 / 0.044
Oleic acid / HMDB00207 / N.D. / N.S. / + / 1.220 / 0.001 / N.D. / N.S.
Palmitic acid / HMDB00220 / N.D. / N.S. / + / 0.821 / 0.018 / N.D. / N.S.
Palmitoleic acid / HMDB03229 / N.D. / N.S. / + / 1.419 / < 0.001 / N.D. / N.S.
Vaccenic acid / HMDB03231 / N.D. / N.S. / + / 1.044 / 0.003 / N.D. / N.S.
Bile & Cholesterols / Cholesterol / HMDB00067 / N.D. / N.S. / + / 0.921 / 0.028 / N.D. / N.S.
Taurocholic acid / HMDB00922 / N.D. / N.S. / – / 1.038 / 0.023 / N.D. / N.S.
Phospholipid metabolism / PC(O-14:0/0:0) / LMGP01060007 / N.D. / N.S. / N.D. / N.S. / – / 2.729 / 0.001
Ethanolamine / HMDB00149 / N.D. / N.S. / – / 0.979 / 0.008 / N.D. / N.S.
Myo-inositol / HMDB00211 / N.D. / N.S. / N.D. / N.S. / + / 1.757 / 0.024
Other lipids / Aminooctanoic acid / HMDB00991 / N.D. / N.S. / N.D. / N.S. / + / 1.515 / 0.034
Glycerol / HMDB00131 / N.D. / N.S. / + / 1.302 / 0.001 / N.D. / N.S.
MG(18:1/0:0/0:0) / HMDB11567 / N.D. / N.S. / + / 0.901 / 0.007 / N.D. / N.S.
Purine metabolism / Arabitol / HMDB01851 / N.D. / N.S. / + / 0.810 / 0.020 / N.D. / N.S.
GMP / HMDB01397 / N.D. / N.S. / + / 1.477 / 0.001 / N.D. / N.S.
Pyrimidine metabolism / Uracil / HMDB00300 / + / 0.850 / 0.040 / N.D. / N.S. / N.D. / N.S.
TCA metabolism / Acetic acid / HMDB00042 / N.D. / N.S. / N.D. / N.S. / + / 1.177 / 0.049
Pyruvic acid / HMDB00243 / N.D. / N.S. / + / 0.969 / 0.008 / N.D. / N.S.
Succinic acid / HMDB00254 / N.D. / N.S. / – / 0.802 / 0.037 / N.D. / N.S.
3-Hydroxybutyric acid / HMDB00357 / + / 1.446 / 0.001 / + / 1.766 / < 0.001 / N.D. / N.S.
Lactic acid / HMDB00190 / – / 1.079 / 0.010 / N.D. / N.S. / N.D. / N.S.
Malic acid / HMDB00156 / N.D. / N.S. / N.D. / N.S. / – / 1.660 / 0.022
Vitamin metabolism / Hydroxyvitamin D3 / N.D. / N.S. / N.D. / N.S. / – / 1.639 / 0.031
Vitamin D3 / HMDB00876 / N.D. / N.S. / N.D. / N.S. / – / 1.801 / 0.025
Other / 3-Hydroxy picolinic acid / HMDB13188 / N.D. / N.S. / + / 1.051 / 0.005 / N.D. / N.S.
Aminomalonic acid / HMDB01147 / N.D. / N.S. / + / 0.899 / 0.017 / N.D. / N.S.
Glucaric acid / HMDB00663 / N.D. / N.S. / + / 1.084 / 0.002 / N.D. / N.S.
Oxalic acid / HMDB02329 / – / 1.778 / < 0.001 / N.D. / N.S. / N.D. / N.S.

Table S-6: Metabolite levels different in the brain of all three strains of mice that swam when compared to their respective controls

Group / Metabolite / ID # / WT / MT1+2KO / MT3KO
Direction / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p) / Direction / Effect size (d) / t-test (p)
Amino acid metabolism / 2-Keto-6-acetamidocaproate / HMDB12150 / N.D. / N.S. / – / 0.870 / 0.036 / N.D. / N.S.
2-Ketobutyric acid / HMDB00005 / N.D. / N.S. / N.D. / N.S. / – / 1.385 / 0.031
2-Pyrrolidone-5-carboxylic acid / HMDB00805 / – / 1.384 / 0.002 / N.D. / N.S. / – / 2.851 / 0.001
Alanine / HMDB00161 / – / 1.271 / 0.004 / – / 0.864 / 0.026 / – / 4.055 / < 0.01
Aspartic acid / HMDB00191 / – / 1.062 / 0.016 / N.D. / N.S. / – / 1.500 / 0.025
Cadaverine / HMDB02322 / – / 1.884 / < 0.01 / N.D. / N.S. / – / 1.789 / 0.006
Cysteine / HMDB00574 / – / 0.992 / 0.022 / N.D. / N.S. / N.D. / N.S.
Glutamine / HMDB00641 / – / 1.244 / 0.003 / N.D. / N.S. / – / 2.254 / 0.003
Glycine / HMDB00123 / N.D. / N.S. / N.D. / N.S. / – / 1.409 / 0.022
Isoleucine / HMDB00172 / – / 1.567 / 0.001 / – / 1.110 / 0.010 / – / 3.191 / < 0.01
Methionine / HMDB00696 / – / 1.246 / 0.007 / – / 0.973 / 0.019 / – / 2.494 / 0.001
Phenylethylamine / HMDB12275 / – / 1.432 / 0.002 / N.D. / N.S. / N.D. / N.S.
Proline / HMDB00162 / – / 1.607 / 0.001 / – / 0.865 / 0.029 / – / 1.404 / 0.036
Serine / HMDB00187 / – / 1.173 / 0.005 / N.D. / N.S. / – / 2.752 / 0.001
Threonine / HMDB00167 / – / 1.120 / 0.012 / N.D. / N.S. / – / 2.065 / 0.005
Valine / HMDB00883 / – / 1.294 / 0.003 / N.D. / N.S. / – / 2.124 / 0.002
Antioxidant metabolism / Pyroglutamic acid / HMDB00267 / – / 1.700 / < 0.01 / N.D. / N.S. / – / 3.224 / < 0.01
Carbohydrate metabolism / Erythrose / HMDB02649 / – / 0.950 / 0.024 / N.D. / N.S. / – / 3.684 / < 0.01
Fructose / HMDB00660 / – / 1.266 / 0.003 / – / 1.802 / < 0.01 / – / 5.008 / < 0.01
Ribose / HMDB00283 / – / 2.080 / < 0.01 / N.D. / N.S. / – / 3.509 / < 0.01
Fucose / HMDB00174 / N.D. / N.S. / – / 0.903 / 0.043 / – / 3.946 / < 0.01
2,3,4-Trihydroxybutyric acid / HMDB00613 / – / 0.973 / 0.014 / – / 0.803 / 0.028 / – / 3.199 / 0.001
Fucose / HMDB00174 / – / 1.164 / 0.003 / N.D. / N.S. / N.D. / N.S.
Gluconic acid / HMDB00625 / – / 1.063 / 0.019 / N.D. / N.S. / N.D. / N.S.
Glucose-6-phosphate / HMDB01401 / N.D. / N.S. / N.D. / N.S. / – / 1.095 / 0.051
Glycerol 3-phosphate / HMDB00126 / N.D. / N.S. / N.D. / N.S. / – / 1.673 / 0.014
Creatine phosphate system / Creatinine / HMDB00562 / N.D. / N.S. / N.D. / N.S. / – / 2.677 / 0.002
Lipid metabolism / Carnitine / HMDB00562 / + / 0.914 / 0.028 / N.D. / N.S. / N.D. / N.S.
Free fatty acids / Arachidonic acid / HMDB01043 / N.D. / N.S. / N.D. / N.S. / – / 2.552 / < 0.01
Eicosanoic acid / HMDB02212 / – / 2.391 / < 0.01 / N.D. / N.S. / – / 4.124 / < 0.01
Glycerol / HMDB00131 / N.D. / N.S. / N.D. / N.S. / – / 5.422 / < 0.01
Heneicosanoic acid / HMDB02345 / – / 0.965 / 0.009 / N.D. / N.S. / – / 3.235 / < 0.01
Margaric acid / HMDB02259 / – / 2.724 / < 0.01 / N.D. / N.S. / – / 5.477 / < 0.01
Nonanoic acid / HMDB00847 / – / 1.295 / 0.005 / N.D. / N.S. / – / 1.806 / 0.009
Oleic acid / HMDB00207 / N.D. / N.S. / N.D. / N.S. / – / 1.210 / 0.031
Palmitic acid / HMDB00220 / N.D. / N.S. / N.D. / N.S. / – / 1.613 / 0.004
Stearic acid / HMDB00827 / N.D. / N.S. / N.D. / N.S. / – / 2.179 / 0.001
Acylglycerides / TG(17:2/20:5/22:4) / LMGL03011744 / N.D. / N.S. / N.D. / N.S. / – / 1.438 / 0.043
2-Monopalmitin / HMDB11533 / N.D. / N.S. / N.D. / N.S. / – / 1.851 / 0.004
Phospholipid metabolism / Ethanolamine / HMDB00149 / – / 0.956 / 0.019 / N.D. / N.S. / N.D. / N.S.
Myo-inositol / HMDB00211 / – / 2.553 / < 0.01 / N.D. / N.S. / – / 4.940 / < 0.01
Phosphocholine / HMDB01565 / N.D. / N.S. / – / 1.145 / 0.006 / N.D. / N.S.
Phosphorylethanolamine / HMDB00224 / N.D. / N.S. / N.D. / N.S. / – / 1.286 / 0.029
Other lipids / Cholesterol / HMDB00067 / N.D. / N.S. / N.D. / N.S. / – / 1.288 / 0.034
Glyceric acid / HMDB06372 / – / 1.023 / 0.020 / N.D. / N.S. / – / 2.130 / 0.004
Glycerol / HMDB00131 / – / 1.699 / < 0.01 / – / 1.085 / 0.018 / N.D. / N.S.
Neurotransmitters / N-acetylaspartate / HMDB00812 / – / 2.243 / < 0.01 / – / 1.071 / 0.006 / – / 3.809 / < 0.01
γ-Aminobutyric acid / HMDB00112 / – / 1.262 / 0.004 / N.D. / N.S. / – / 2.834 / 0.001
Purine metabolism / Adenine / HMDB00034 / – / 1.345 / < 0.01 / N.D. / N.S. / – / 1.154 / 0.030
AMP / HMDB00045 / – / 1.084 / 0.016 / N.D. / N.S. / – / 1.583 / 0.025
Guanidylic acid (GMP) / HMDB01397 / N.D. / N.S. / N.D. / N.S. / + / 1.155 / 0.020
Inosine / HMDB00195 / – / 1.364 / 0.002 / N.D. / N.S. / – / 1.759 / 0.019
Myo-Inositol phosphate / HMDB00213 / N.D. / N.S. / N.D. / N.S. / – / 1.646 / 0.012
Ribitol / HMDB00508 / – / 1.446 / 0.002 / N.D. / N.S. / – / 2.279 / 0.004
Ribose-5-phosphate / HMDB01548 / – / 1.201 / 0.009 / N.D. / N.S. / – / 2.514 / 0.001
Pyrimidine metabolism / Cytosine / HMDB00630 / N.D. / N.S. / – / 0.902 / 0.019 / – / 2.028 / 0.010
Uracil / HMDB00300 / N.D. / N.S. / – / 1.274 / 0.003 / N.D. / N.S.
Uridine / HMDB00296 / N.D. / N.S. / N.D. / N.S. / – / 1.356 / 0.041
TCA metabolism / 2-Ketoglutaric acid / HMDB00208 / – / 1.153 / 0.004 / N.D. / N.S. / – / 1.363 / 0.048
Citric acid / HMDB00094 / – / 1.242 / 0.006 / N.D. / N.S. / – / 2.717 / 0.001
Fumaric acid / HMDB00134 / – / 2.104 / < 0.01 / N.D. / N.S. / – / 5.924 / < 0.01
Lactic acid / HMDB00190 / – / 2.037 / < 0.01 / N.D. / N.S. / – / 3.273 / < 0.01
Malic acid / HMDB00156 / – / 1.783 / < 0.01 / – / 0.760 / 0.042 / – / 2.951 / < 0.01
Succinic acid / HMDB00254 / N.D. / N.S. / N.D. / N.S. / – / 1.520 / 0.018
Other / 3-Hydroxy picolinic acid / HMDB13188 / N.D. / N.S. / N.D. / N.S. / – / 1.730 / 0.009
Aminomalonic acid / HMDB01147 / – / 1.216 / 0.006 / N.D. / N.S. / – / 1.974 / 0.006
Gluconic acid / HMDB00625 / N.D. / N.S. / N.D. / N.S. / – / 3.483 / < 0.01
Oxalic acid / HMDB02329 / – / 1.525 / 0.001 / N.D. / N.S. / – / 3.160 / < 0.01

Additional figures

Fig.S-3: Gastrocnemius and circulating metabolites that differed significantly between the knockout and wild-type mice during unchallenged conditions. The metabolites that differed between the MT1+2KO and WT or MT3KO and WT mice are shown in bold. They are illustrated within the potentially affected metabolic pathways. Their respective ANOVA box-and-whisker plots are also shown and linked to the respective metabolites through a perforated line. The relative, transformed (scaled) abundance of each metabolite is indicated on the y-axis. The legend for the box-and-whisker plots (x-axis) are shown in the upper left corner. Dark gray box, MT1+2KO; Open/white box, MT3KO; Light grey box, WT; FFA, free fatty acids; MG, monoacylglycerol.

Fig. S-4: Hepatic metabolites that differed significantly between the knockout and wild-type mice during unchallenged conditions.

Fig.S-5: Cerebral metabolites that differed significantly between the knockout and wild-type mice during unchallenged conditions.

Fig.S-6: Gastrocnemius and circulating metabolites that differed significantly between the knockout and wild-type mice during challenged conditions. FFA, free fatty acids; MG, monoacylglycerol; PC, Phosphatidylchonline; PE, Phosphattidylethanolamine

Fig.S-7: Hepatic metabolites that differed significantly between the knockout and wild-type mice during challenged conditions.

Fig. S-8: Cerebral metabolites that differed significantly between the knockout and wild-type mice after one hour swim. The metabolites that differed between the MT1+2KO and WT or MT3KO and WT mice are shown in bold. They are illustrated within the potentially affected metabolic pathways with the top figure (A) focussing on the central carbon and amino acid metabolism and bottom figure (B) showing the lipids and fatty acids. Dark gray box, MT1+2KO; Open/white box, MT3KO; Light grey box, WT; FFA, free fatty acids; 2K6AACA, 2-keto-6-acetamidocaproic acid; 2P5CA, 2-pyrrolidone-5-carboxylic acid.