Vitamin B6 deficiency during pregnancy translates into a severe vitamin B6 deficiency (plasma levels decreased by 97%) in new-born rats. Further, hallmarks are increased (+89%) concentrations of homocysteine, gross changes in gene methylation and expression, and metabolic alterations including lipid metabolism. This study focuses on determining the effects of vitamin B6-deficiency on cardiolipin composition and oxidative phosphorylation in liver. For this purpose, hepatic cardiolipin composition was analyzed by means of LC/MS/MS, and mitochondrial oxygen consumption was determined by using a Clark-type electrode in a rat model of vitamin B6 deficiency. Liver mitochondria from new-born rats with pre-term vitamin B6 deficiency responded with substantial alterations in cardiolipin composition that include the following changes in the amounts of cardiolipin incorporated fatty acids: increase in C16, decrease in C18, decrease in saturated fatty acid, as well as increase in amount of oxidized cardiolipin species. These changes were accompanied by significantly decreased capacity of oxidative phosphorylation. In conclusion, vitamin B6 deficiency in new born rats induces massive alterations of cardiolipin composition and function of liver mitochondria. These findings support the importance of sufficient periconceptional supply of vitamin B6 to prevent vitamin B6 deficiency.
Impact statement
Vitamin B6 (VitB6) is an active co-enzyme for more than 150 enzymes and is required for a great diversity of biosynthesis and metabolic reactions. There is an increased need for VitB6 during pregnancy and sufficient supply of VitB6 is crucial for the prevention of cleft palate and neural tube defects. We show that liver mitochondria from new-born rats with pre-term VitB6 deficiency respond with substantial alterations in cardiolipin (CL) composition and in the amount of oxidized CL species. These changes are associated with a decrease in the efficiency of oxidative phosphorylation. The results of this study support the significance of sufficient supply of VitB6 during pregnancy (and periconceptional) for diminishing the number of early abortions and minimizing malformation. The established link between VitB6 deficiency, CL composition, and mitochondrial respiration/energy production provides mechanistic insight as to how the VitB6 deficiency translates into the known pathophysiological and clinically relevant conditions.
ParraMStahlSHellmannH.Vitamin B6 and its role in cell metabolism and physiology. Cells2018;
7: pii:E84
2.
UelandPMUlvikARios-AvilaLMidttunØGregoryJF.Direct and functional biomarkers of vitamin B6 status. Annu Rev Nutr2015;
35:33–70
3.
BenderDA.Non-nutritional uses of vitamin B6. Br J Nutr1999;
81:7–20
4.
BordoniACabriniLMarchettiMDanesiFBochicchioDBiagiPLMaranesiM.Vitamin B6 deficiency and dietary fats: effects on lipid composition and glutathione peroxidase activity in rat liver. Ann Nutr Metab2006;
50:305–12
5.
PantonKKFarupPGSagenESirumUFAsbergA.Vitamin B6 in plasma – sample stability and the reference limits. Scand J Clin Lab Invest2013;
73:476–9
6.
CortesFMelladoCPardoRAVillarroelLAHertrampfE.Wheat flour fortification with folic acid: changes in neural tube defects rates in Chile. Am J Med Genet2012;
158A:1885–90
7.
ElmazarMMThielRNauH.Effect of supplementation with folinic acid, vitamin B6, and vitamin B12 on valproic acid-induced teratogenesis in mice. Fundam Appl Toxicol1992;
18:389–94
8.
JacobssonCGranströmG.Clinical appearance of spontaneous and induced first and second branchial arch syndromes. Scand J Plast Reconstr Surg Hand Surg1997;
31:125–36
9.
KrapelsIPvan RooijIAOckeMCvan CleefBAKuijpers-JagtmanAMSteegers-TheunissenRP.Maternal dietary B vitamin intake, other than folate, and the association with orofacial cleft in the offspring. Eur J Nutr2004;
43:7–14
10.
SmolinLABenevengaNJ.Factors affecting the accumulation of homocyst(e)ine in rats deficient in vitamin B-6. J Nutr1984;
114:103–11
11.
CoppAJ.Prevention of neural tube defects: vitamins, enzymes and genes. Curr Opin Neurol1998;
11:97–102
12.
EskesTK.Folates and the fetus. Eur J Obstet Gynecol Reprod Biol1997;
71:105–11
13.
HolFAvan der PutNMGeurdsMPHeilSGTrijbelsFJHamelBCMarimanECBlomHJ.Molecular genetic analysis of the gene encoding the trifunctional enzyme MTHFD (methylenetetrahydrofolate-dehydrogenase, methenyltetrahydrofolate-cyclohydrolase, formyltetrahydrofolate synthetase) in patients with neural tube defects. Clin Genet1998;
53:119–25
14.
PietrzikKPrinz-LangenohlRThorandB.Mikronährstoffe in der schwangerschaft. Z Geburtshilfe Neonatol1997;
201:21–4
15.
Steegers-TheunissenRPSmithellsRWEskesTK.Update of new risk factors and prevention of neural-tube defects. Obstet Gynecol Surv1993;
48:287–93
16.
EskesTK.Open or closed? a world of difference: a history of homocysteine research. Nutrit Revs1998;
56:236–44
17.
Goddijn-WesselTAWoutersMGvan de MolenEFSpuijbroekMDSteegers-TheunissenRPBlomHJBoersGHEskesTK.Hyperhomocysteinemia: a risk factor for placental abruption or infarction. Eur J Obstet Gynecol Reprod Biol1996;
66:23–9
18.
WeingärtnerJFanghänelJBienengräberVGundlachK.Initial findings on teratological and developmental relationships and differences between neural tube defects and facial clefting. First experimental results. J Craniomaxillofacial Surg2005;
33:297–300
19.
DepeintFBruceWRShangariNMehtaRO'BrienPJ.Mitochondrial function and toxicity: role of the B vitamin family on mitochondrial energy metabolism. Chem Biol Interact2006;
163:94–112
20.
DasarathyJGrucaLLBennettCParimiPSDuenasCMarczewskiSFierroJLKalhanSC.Methionine metabolism in human pregnancy. Am J Clin Nutr2010;
91:357–65
21.
CabriniLBochicchioDBordoniASassiSMarchettiMMaranesiM.Correlation between dietary polyunsaturated fatty acids and plasma homocysteine concentration in vitamin B6-deficient rats. Nutr Metab Cardiovasc Dis2005;
15:94–9
22.
McNeilCJBeattieJHGordonMJPirieLPDuthieSJ.Nutritional B vitamin deficiency disrupts lipid metabolism causing accumulation of proatherogenic lipoproteins in the aorta adventitia of ApoE null mice. Mol Nutr Food Res2012;
56:1122–30
23.
ZhaoMRalatMAda SilvaVGarrettTJMelnykSJamesSJGregoryJF.3rd.Vitamin B-6 restriction impairs fatty acid synthesis in cultured human hepatoma (HepG2) cells. Am J Physiol Endocrinol Metab2013;
304:E342–51
24.
KaimotoTShibuyaMNishikawaKMaedaH.High incidence of lipid deposition in the liver of rats fed a diet supplemented with branched-chain amino acids under vitamin B6 deficiency. J Nutr Sci Vitaminol2013;
59:73–8
25.
NogaAAVanceDE.Insights into the requirement of phosphatidylcholine synthesis for liver function in mice. J Lipid Res2003;
44:1998–2005
26.
VanceDE.Role of phosphatidylcholine biosynthesis in the regulation of lipoprotein homeostasis. Curr Opin Lipidol2008;
19:229–34
27.
SchlameMGreenbergML.Biosynthesis, remodeling and turnover of mitochondrial cardiolipin. Biochim Biophys Acta2017;
1862:3–7
28.
NeusteinHBLuriePRDahmsBTakahashiM.An X-linked recessive cardiomyopathy with abnormal mitochondria. Pediatrics1979;
64:24–9
29.
AcehanDXuYStokesDLSchlameM.Comparison of lymphoblast mitochondria from normal subjects and patients with Barth syndrome using electron microscopic tomography. Lab Invest2007;
87:40–8
30.
FrohmanMA.Role of mitochondrial lipids in guiding fission and fusion. J Mol Med2015;
3:263–9
31.
GürtlerSOttoOHeiseNScholzFLaporteAElsnerMJörnsAWeinertSWolkeCDöringMJansingSGardemannALendeckelUSchildL.Tafazzin-dependent cardiolipin composition in C6 glioma cells correlates with changes in mitochondrial and cellular functions, and cellular proliferation. Biochim Biophys Acta Mol Cell Biol Lipids2019;
1864:452–65
32.
ParadiesGParadiesVDe BenedictisVRuggieroFMPetrosilloG.Functional role of cardiolipin in mitochondrial bioenergetics. Biochim Biophys Acta2014;
1837:408–17
33.
AcehanDMalhotraAXuYRenMStokesDLSchlameM.Cardiolipin affects the supramolecular organization of ATP synthase in mitochondria.Biophys J2011;
100:2184–92
KiebishMAHanXChengHChuangJHSeyfriedTN.Cardiolipin and electron transport chain abnormalities in mouse brain tumor mitochondria: lipidomic evidence supporting the Warburg theory of cancer. J Lipid Res2008;
49:2545–56
36.
RajaVGreenbergML.The functions of cardiolipin in cellular metabolism-potential modifiers of the Barth syndrome phenotype. Chem Phys Lipids2014;
179:49–56
37.
SchlameM.Cardiolipin remodeling and the function of tafazzin. Biochim Biophys Acta2013;
1831:582–8
38.
DudekJChengIFBalleiningerMVazFMStreckfuss-BömekeKHübscherDVukoticMWandersRJRehlingPGuanK.Cardiolipin deficiency affects respiratory chain function and organization in an induced pluripotent stem cell model of Barth syndrome. Stem Cell Res2013;
11:806–19
39.
FuertigRCeciACamusSMBezardELuippoldAHHengererB.LC-MS/MS-based quantification of kynurenine metabolites, tryptophan, monoamines and neopterin in plasma, cerebrospinal fluid and brain. Bioanalysis2016;
8:1903–17
40.
OhligTLeDVGardemannAWolkeCGürtlerSPeterDSchildLLendeckelU.Effects of siRNA-dependent knock-down of cardiolipin synthase and tafazzin on mitochondria and proliferation of glioma cells. Biochim Biophys Acta2018;
1863:379–87
41.
Nourooz-ZadehJLiuEHYhlenBAngardEEHalliwellB.F4-Isoprostanes as specific marker of docosahexaenoic acid peroxidation in Alzheimer´s disease. J Neurochem1999;
72:734–40
42.
BradfordMM.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt Biochem1976;
72:248–54
43.
ChilukotiRKLendeckelJDarmKBukowskaAGoetteASühlingMUtpatelKPetersBHomuthGVölkerUWolkeCScharfCLendeckelU.Integration of “omics” techniques: dronedarone affects cardiac remodeling in the infarction border zone. Exp Biol Med2018;
243:895–910
44.
VanderlindeRE.Review of pyridoxal phosphate and the transaminases in liver disease. Ann Clin Lab Sci1986;
16:79–93
45.
SchulmanMPRichertDA.Heme synthesis in vitamin B6 and pantothenic acid deficiencies. J Biol Chem1957;
226:181–9
46.
WeissbachHBogdanskiDFRedfieldBGUdenfriendS.Studies on the effect of vitamin B6 on 5-hydroxytryptamine (serotonin) formation. J Biol Chem1957;
227:617–24
47.
FerroYCarèIMazzaEProvenzanoFColicaCTortiCRomeoSPujiaAMontalciniT.Protein and vitamin B6 intake are associated with liver steatosis assessed by transient elastography, especially in obese individuals. Clin Mol Hepatol2017;
23:249–59
48.
BhateAParkerDJBebeeTWAhnJArifWRashanEHChorghadeSChauALeeJHAnakkSCarstensRPXiaoXKalsotraA.ESRP2 controls an adult splicing programme in hepatocytes to support postnatal liver maturation. Nat Commun2015;
6:8768
49.
LiuZLiPZhaoZHZhangYMaZMWangSX.Vitamin B6 prevents endothelial dysfunction, insulin resistance, and hepatic lipid accumulation in apoe (-/-) mice fed with High-Fat diet. J Diabetes Res2016;
2016:1748065
50.
SchildLLendeckelUGardemannAWiswedelISchmidtCAWolkeCWaltherRGrabarczykPBusemannC.Composition of molecular cardiolipin species correlates with proliferation of lymphocytes. Exp Biol Med2012;
237:372–9
51.
HartzJWMortonREWaiteMMMorrisHP.Correlation of fatty acyl composition of mitochondrial and microsomal phospholipid with growth rate of rat hepatomas. Lab Invest1982;
46:73–8
52.
DevlinAMSinghRWadeREInnisSMBottiglieriTLentzSR.Hypermethylation of Fads2 and altered hepatic fatty acid and phospholipid metabolism in mice with hyperhomocysteinemia. J Biol Chem2007;
282:37082–90
53.
ZhaoMLamersYRalatMACoatsBSChiYYMullerKEBainJRShankarMNNewgardCBStacpoolePWGregoryJF3rd.Marginal vitamin B-6 deficiency decreases plasma (n-3) and (n-6) PUFA concentrations in healthy men and women. J Nutr2012;
142:1791–7
54.
CostaRHKalinichenkoVVHoltermanAXWangX.Transcription factors in liver development, differentiation, and regeneration. Hepatology2003;
38:1331–47
55.
KyrmiziIHatzisPKatrakiliNTroncheFGonzalezFJTalianidisI.Plasticity and expanding complexity of the hepatic transcription factor network during liver development. Genes Dev2006;
20:2293–305
56.
IkonNRyanRO.Cardiolipin and mitochondrial cristae organization. Biochim Biophys Acta Biomembr2017;
1859:1156–63
57.
SchlameMRenM.The role of cardiolipin in the structural organization of mitochondrial membranes. Biochim Biophys Acta2009;
1788:2080–3
58.
ClaypoolSMBoontheungPMcCafferyJMLooJAKoehlerCM.The cardiolipin transacylase, tafazzin, associates with two distinct respiratory components providing insight into Barth syndrome. MBoC2008;
19:5143–55
