Muscle and serum acylcarnitine profiles in dairy cows during the periparturient period
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1 Muscle nd serum cylcrnitine profiles in diry cows during the periprturient period Y. Yng, 1 C. Prehn, 2 J. Admski, 2 J. Rehge, 3 S. Dänicke, 4 H. Suerwein, 1 H. Sdri 1 1 Institute of Animl Science, Physiology & Hygiene Unit, University of Bonn, Germny 2 Institute of Experimentl Genetics, Genome Anlysis Center, Helmholtz Zentrum München, Germn Reserch Center for Environmentl Helth, Neuhererg, Germny 3 Clinic for Cttle, University of Veterinry Medicine, Hnnover, Germny 4 Institute of Animl Nutrition, Friedrich-Loeffler-Institute, Brunschweig, Germny 67 th Annul Meeting of the Europen Federtion of Animl Science, Belfst, UK, 29 Aug 2 Sept 2016
2 Introduction Negtive energy lnce nd relese of free ftty cids into the circultion during erly lcttion Turning skeletl muscle into the mjor site for use of ft-derived fuels Excess lipid supply nd susequent lipid ccumultion in insulin-sensitive tissues (Vn Epps-Fung et l., 1997; Pires et l., 2007; Schoonemn et l., 2013) 67 th Annul Meeting of the Europen Federtion of Animl Science, Belfst, UK, 29 Aug 2 Sept
3 The Crnitine Shuttle TCA Adpted from: 3
4 Ojective of the Study To chrcterize the concentrtions of crnitine nd its cyl esters (cylcrnitines; ACC) oth in lood serum nd in skeletl muscle of diry cows to ddress potentil chnges in the cpcity for mitochondril β-oxidtion of ftty cids in skeletl muscle in context with the negtive energy lnce typicl for erly lcttion. 67 th Annul Meeting of the Europen Federtion of Animl Science, Belfst, UK, 29 Aug 2 Sept
5 Mterils nd Methods 11 Pluriprous Germn Holstein cows Biopsies from M. semitendinosus nd lood smpling: dy -21, 1, 21, nd 70 reltive to clving Muscle nd serum ACC profiles: y FIA-MS/MS profiling through trgeted metolomics pproch (AsoluteIDQ p180 Kit; Biocrtes Life Sciences AG, Innsruck, Austri). 67 th Annul Meeting of the Europen Federtion of Animl Science, Belfst, UK, 29 Aug 2 Sept
6 Sttisticl Anlysis Mterils nd Methods The PROC MIXED procedure of SAS (9.3) using repeted mesures: - Fixed effect: Time (smpling dy) - Rndom effect: Cow Correltion nlysis using PROC CORR The threshold of significnce: P < 0.05; Trends: 0.05 < P < 0.10 Dt presented s mens ± SEM 67 th Annul Meeting of the Europen Federtion of Animl Science, Belfst, UK, 29 Aug 2 Sept
7 μm μm μm Crnitine Serum short chin ACC C2 (Acetylcrnitine) 3 2 Serum medium chin ACC C7-DC (Pimelylcrnitine) C3 (Propionylcrnitine) C4 (Butyrylcrnitine) C8 (Cprylylcrnitine) C4:1 (Butenoylcrnitine) C5 (Isovlerylcrnitine) Dys reltive to clving Dys reltive to clving P < 0.05; except C4:1 nd C5 (P < 0.10 )
8 μm μm Serum long chin ACC C14:2 (Tetrdecdienoylcrnitine) C16 (Plmitoylcrnitine) C16-OH (Hydroxyplmitoylcrnitine) C16:2 (Hexdecdienoylcrnitine) C18 (Sterylcrnitine) C18:1-OH (Hydroxyoleylcrnitine) Dys reltive to clving P < 0.02; except C14:2, C16-OH, nd C16:2 (P < 0.10 )
9 pmol/mg tissue pmol/mg tissue Crnitine Muscle short chin ACC C2 (Acetylcrnitine) Muscle medium chin ACC 1.2 C6 (Hexnoylcrnitine) C3 (Propionylcrnitine) 0 4 C4-OH (Mlonylcrnitine) c C6:1 (Hexenoylcrnitine) C5-OH (Hydroxyvlerylcrnitine) c C5:1 (3-Methyl-crotonylcrnitine) Dys reltive to clving 0.0 Dys reltive to clving P < 0.03; except crnitine, C2, nd C6: non significnt
10 pmol/mg tissue Muscle long chin ACC C14 (Myristylcrnitine) C16 (Plmitoylcrnitine) C16:1 (Plmitoleylcrnitine) c C18 (Sterylcrnitine) C18:1 (Oleylcrnitine) C18:2 (Linoleylcrnitine) c 0 Dys reltive to clving P < 0.001; except C14 (P < 0.09 )
11 Metolites correlting with NEFA (cross ll time points) Serum Muscle C 1 4 : 1 C 1 6 -O H C 1 6 : 2 C 6 C 1 4 : 2 M C A C C C 4 -O H C 7 -D C C 1 6 L C A C C C 8 S C A C C C 2 C 1 8 : 1 C 1 8 M C A C C C 1 8 C 5 : 1 C 1 6 : 1 C 6 : 1 C 4 -O H L C A C C C 1 8 : 1 C Person correltion coefficients P < 0.04; except Serum C16-OH (P < 0.07)
12 CPT-I Rtio (crnitine/c16+c18)* Serum P < Muscle Dys reltive clving *Ppstel et l. (2016). IJC Metolic & Endocrine. 11:13 18.
13 CPT-II Rtio (C16+C18:1/C2)* Serum P < Muscle c c c c Dys reltive clving *Gempel et l., J. Inherit. Met. Dis. 25:17-27.
14 Conclusions Serum concentrtions of crnitine decresed with the onset of lcttion, proly due to incresed crnitine excretion in milk nd incresed crnitine uptke y the muscle to mintin the intrcellulr concentrtions. ACC profiles demonstrted incresed lipid flux nd consequently incomplete or overloded ftty cids oxidtion in the skeletl muscle round clving. These dt suggest tht post-cpt1 events including deficiency or impired CPT2 function nd/or depletion of severl TCA cycle intermedites re more prole fctors cusing ccumultion of long chin ACC in the muscle round prturition. 67 th Annul Meeting of the Europen Federtion of Animl Science, Belfst, UK, 29 Aug 2 Sept
15 Thnk you for your ttention 67 th Annul Meeting of the Europen Federtion of Animl Science, Belfst, UK, 29 Aug 2 Sept 2016
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18 Metolites correlting with RQUICKI (cross ll time points) M u s c le C 1 6 M u s c le C 1 8 : 1 M u s c le C 1 6 : 1 S e ru m C 7 -D C S e ru m C 1 4 : 2 S e ru m C 6 S e ru m C 8 S e ru m M C A C C Correltion coefficients P < 0.04; except serum C7-DC nd muscle C16, C18:1, nd C16:1 (P < 0.09)
19 R Q U IC K I, d M us c le C 1 8 M us c le M C A C M us c le C S e rum C M us c le C S e rum C 5 S e rum C 3 S e rum C 1 8 S e rum M C A C Ins ulin S e rum C 2 S e rum S C A C C o r r e l tio n c o e ffic ie n ts P < 0.04, except Muscle C16.1, C18.2, MCAC, C18, nd Serum C14.1 (P = , , , , nd , respectively) 19
20 μ m o l/l N E F A P < D y s re l tiv e to c lv in g 20
21 Acylcrnitine BC code Anlyte C0 Crnitine (free) C2 Acetylcrnitine C3 Propionylcrnitine C3:1 Propenoylcrnitine C3-OH Hydroxypropionylcrnitine C4 Butyrylcrnitine / Isoutyrylcrnitine C4:1 Butenoylcrnitine C4-OH (C3-DC) Hydroxyutyrylcrnitine (Mlonylcrnitine) C5 Isovlerylcrnitine / 2-Methylutyrylcrnitine / Vlerylcrnitine C5:1 Tiglylcrnitine / 3-Methyl-crotonylcrnitine C5:1-DC Glutconylcrnitine / Mesconylcrnitine C5-DC (C6-OH) Glutrylcrnitine (Hydroxyhexnoylcrnitine [= Hydroxycproylcrnitine]) C5-M-DC Methylglutrylcrnitine C5-OH (C3-DC-M) Hydroxyisovlerylcrnitine / Hydroxy-2-methylutyryl / Hydroxyvlerylcrnitine (Methylmlonylcrnitine) C6 (C4:1-DC) Hexnoylcrnitine [= Cproylcrnitine] (Fumrylcrnitine) C6:1 Hexenoylcrnitine C7-DC Pimelylcrnitine C8 Octnoylcrnitine [= Cprylylcrnitine] C9 Nonnoylcrnitine [= Pelrgonylcrnitine] C10 Decnoylcrnitine [= Cprylcrnitine] C10:1 Decenoylcrnitine C10:2 Decdienoylcrnitine C12 Dodecnoylcrnitine [= Lurylcrnitine] C12:1 Dodecenoylcrnitine C12-DC Dodecnedioylcrnitine C14 Tetrdecnoylcrnitine [= Myristylcrnitine] C14:1 Tetrdecenoylcrnitine [= Myristoleylcrnitine] C14:1-OH Hydroxytetrdecenoylcrnitine [= Hydroxymyristoleylcrnitine] C14:2 Tetrdecdienoylcrnitine C14:2-OH Hydroxytetrdecdienoylcrnitine C16 Hexdecnoylcrnitine [= Plmitoylcrnitine] C16:1 Hexdecenoylcrnitine [= Plmitoleylcrnitine] C16:1-OH Hydroxyhexdecenoylcrnitine [= Hydroxyplmitoleylcrnitine] C16:2 Hexdecdienoylcrnitine C16:2-OH Hydroxyhexdecdienoylcrnitine C16-OH Hydroxyhexdecnolycrnitine [= Hydroxyplmitoylcrnitine] C18 Octdecnoylcrnitine [= Sterylcrnitine] C18:1 Octdecenoylcrnitine [= Oleylcrnitine] C18:1-OH Hydroxyoctdecenoylcrnitine [= Hydroxyoleylcrnitine] C18:2 Octdecdienoylcrnitine [= Linoleylcrnitine] 21
22 Muscle Acylcrnitines Vlid dt (%) Serum Acylcrnitines Vlid dt (%) C0 95.5% C0 52% C2 95.5% C2 100% C3 95.5% C3 100% C5-OH (C3-DC-M) 95.5% C5-OH (C3-DC-M) 0% C3-OH 0.0% C3-OH 55% C3:1 0.0% C3:1 0% C4 95.5% C4 100% C3-DC (C4-OH) 95.5% C3-DC (C4-OH) 61% C4:1 25.0% C4:1 43% C5 95.5% C5 100% C5-DC (C6-OH) 43.2% C5-DC (C6-OH) 0% C5-M-DC 0.0% C5-M-DC 0% C5:1 86.4% C5:1 84% C5:1-DC 34.1% C5:1-DC 77% C6 (C4:1-DC) 90.9% C6 (C4:1-DC) 80% C6:1 86.4% C6:1 30% C7-DC 22.7% C7-DC 75% C8 2.3% C8 80% C9 0.0% C9 25% C10 2.3% C10 57% C10:1 0.0% C10:1 0% C10:2 0.0% C10:2 0% C12 6.8% C12 9% C12-DC 0.0% C12-DC 0% C12:1 0.0% C12:1 0% C % C14 50% C14:1 22.7% C14:1 98% C14:1-OH 25.0% C14:1-OH 64% C14:2 2.3% C14:2 80% C14:2-OH 2.3% C14:2-OH 27% C % C16 89% C16-OH 2.3% C16-OH 75% C16:1 81.8% C16:1 66% C16:1-OH 27.3% C16:1-OH 64% C16:2 11.4% C16:2 77% C16:2-OH 4.5% C16:2-OH 48% C % C18 95% C18:1 93.2% C18:1 70% C18:1-OH 2.3% C18:1-OH 50% C18:2 93.2% C18:2 50% 22
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