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Integrating RNA-sequencing and untargeted LC–MS metabolomics to evaluate the effect of lysine deficiency on hepatic functions in Holstein calves

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Abstract

Lysine (Lys) is majorly metabolized in the liver. The liver functional consequences of a dietary Lys deficiency in young Holstein calves are unknown. This study aimed to investigate the effects of Lys deficiency in Holstein calf livers using RNA-sequencing and untargeted LC–MS metabolomics. Calves (n = 36; initial body weight 101.2 ± 10.8 kg; 90-day-old) were fed restricted diets, for 90 days, containing 19.2% crude protein that varied in Lys content (PC group 1.21%; PC-Lys group 0.85%; dry matter basis) for 90 days. Body weight, average daily gain, gain/feed, and Lys intake were significantly decreased in response to Lys deficiency (P < 0.05). Dry matter intake was not altered (P > 0.05). Network and pathway analyses revealed that noradrenaline, adenosine 5′-monophosphate, acetyl-CoA, and coenzyme A were significantly decreased. Regulating of lipolysis in adipocytes pathway and fatty acid degradation pathway were downregulated. We also identified eight significantly differentially expressed genes (SDEGs), among which adrenoceptor beta 2 (ADRB2), WAP four-disulfide core domain 2 (WFDC2), and claudin-4 (CLDN4) were associated with inhibition of lipolysis, and carbon catabolite repression 4-like (CCRN4L), FOS like 2 (FOSL2), and arginase 2 (ARG2) were associated with inhibiting lipid synthesis. Correlation tests showed that coenzyme A was strongly correlated with SDEGs (0.82 ≤|r|≤ 0.96). Acetyl-CoA and adenosine 5′-monophosphate were strongly correlated with CCRN4L (0.90 ≤|r|≤ 0.92), indicating a strong correlation between the changes in SDEGs and these metabolites. In conclusion, Lys deficiency caused dysplasia and affected lipid metabolism in the liver by inhibiting lipolysis and lipid synthesis in calves.

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References

  • Abe M, Iriki T, Funaba M, Onda S (1998) Limiting amino acids for a corn and soybean meal diet in weaned calves less than three months of age. J Anim Sci 76:628–636

    Article  CAS  PubMed  Google Scholar 

  • Alam MR, Yoshizawa F, Sugahara K (2012) Oral administration of lysine restores food intake and ventromedial hypothalamic dopamine in chicken on a lysine-free diet. British Poult Sci 53:651–657

    Article  CAS  Google Scholar 

  • Ameen M, Palmer T, Oberholzer VG (1987) Inhibition of bovine liver lysine-ketoglutarate reductase by urea cycle metabolites and saccharine. Biochem Int 14:589–595

    CAS  PubMed  Google Scholar 

  • Anja V, Juliane B, Masataka N, Sandra H, Ulrich B, Wolfgang M (2015) Transsynaptic tracing from taste receptor cells reveals local taste receptor gene expression in gustatory ganglia and brain. J Neurosci 35:9717–9729

    Article  CAS  Google Scholar 

  • Association of Official Analytical Chemists (2012) Official methods of analysis of the association of offical analytical chemists. AOAC International, Gaithersburg

    Google Scholar 

  • Bos JL (2006) Epac proteins: multi-purpose cAMP targets. Trends Biochem Sci 31:680–686

    Article  CAS  PubMed  Google Scholar 

  • Carrasco R, Arrizon AA, Plascencia A, Torrentera NG, Zinn RA (2013) Comparative feeding value of distillers dried grains plus solubles as a partial replacement for steam-flaked corn in diets for calf-fed Holstein steers: characteristics of digestion, growth performance, and dietary energetics. J Anim Sci 91:1801–1810

    Article  CAS  PubMed  Google Scholar 

  • Ci̇Velek T, Bi̇Rdane F, Kabu M, Cingi CC, Acar A (2013) Effects of methionine and lysine on metabolic profile in dairy cattle during periparturient period. Kafkas Univ Vet Fak 19:423–432

    Google Scholar 

  • Dann HM, Drackley JK (2005) Carnitine palmitoyltransferase I in liver of periparturient dairy cows: effects of prepartum intake, postpartum induction of ketosis, and periparturient disorders. J Dairy Sci 88:3851–3859

    Article  CAS  PubMed  Google Scholar 

  • Donahue PB, Schwab CG, Quigley JD, Hylton WE (1985) Methionine deficiency in early-weaned dairy calves fed pelleted rations based on corn and alfalfa or corn and soybean proteins. J Dairy Sci 68:681–693

    Article  CAS  PubMed  Google Scholar 

  • Dunn WA, Rettura G, Seifter E, Englard S (1984) Carnitine biosynthesis from gamma-butyrobetaine and from exogenous protein-bound 6-N-trimethyl-l-lysine by the perfused guinea pig liver. Effect of ascorbate deficiency on the in situ activity of gamma-butyrobetaine hydroxylase. J Biol Chem 259:10764–10770

    CAS  PubMed  Google Scholar 

  • Flaga J, Korytkowski Å, GãRka P, Kowalski ZM (2018) Age-related changes in mRNA expression of selected surface receptors in lymphocytes of dairy calves. Pol J Vet Sci 21:213–216

    CAS  PubMed  Google Scholar 

  • Friis UG, Madsen K, Stubbe J, Hansen PB, Svenningsen P, Bie P, Skott O, Jensen B (2013) Regulation of renin secretion by renal juxtaglomerular cells. Pflügers Arch Eur J Physiol 465:25–37

    Article  CAS  Google Scholar 

  • Green CB, Douris N, Kojima S, Strayer CA, Fogerty J, Lourim D, Keller SR, Besharse JC (2007) Loss of Nocturnin, a circadian deadenylase, confers resistance to hepatic steatosis and diet-induced obesity. Proc Natl Acad Sci USA 104:9888–9893

    Article  CAS  PubMed  Google Scholar 

  • Grummer RR (1993) Etiology of lipid-related metabolic disorders in periparturient dairy cows. J Dairy Sci 76:3882–3896

    Article  CAS  PubMed  Google Scholar 

  • Halls ML, Cooper DMF (2011) Regulation by Ca2+-signaling pathways of adenylyl cyclases. CSH Perspect Biol 3:1–22

    CAS  Google Scholar 

  • Hee SW, Tsai SH, Chang YC, Chang CJ, Yu IS, Lee PC, Lee WJ, Chang EYC, Chuang LM (2012) The Role of Nocturnin in Early Adipogenesis and Modulation of Systemic Insulin Resistance in Human. Obesity 20:1558–1565

    Article  CAS  PubMed  Google Scholar 

  • Hiroyuki K, Aiko I, Akane A, Kazuki N, Masaya K (2012) Effects of dietary low level of threonine and lysine on the accumulation of intramuscular fat in porcine muscle. Biosci Biotechnol Biochem 76:2347–2350

    Article  CAS  Google Scholar 

  • Katsumata M, Kyoya T, Ishida A, Ohtsuka M, Nakashima K (2012) Dose-dependent response of intramuscular fat accumulation in longissimus dorsi muscle of finishing pigs to dietary lysine levels. Livest Sci 149:41–45

    Article  Google Scholar 

  • Keisuke M, Fumika Y, Masanari T, Masakazu I (2014) Effects of dietary lysine/protein ratio and fat levels on growth performance and meat quality of finishing pigs. Anim Sci J 85:427–434

    Article  CAS  Google Scholar 

  • Keogh K, Kenny DA, Cormican P, Kelly AK, Waters SM (2016) Effect of dietary restriction and subsequent re-alimentation on the transcriptional profile of hepatic tissue in cattle. BMC Genom 17:244

    Article  CAS  Google Scholar 

  • Kim J, Lee KS, Kwon DH, Bong JJ, Jeong JY, Nam YS, Lee MS, Liu X, Biak M (2014) Severe dietary lysine restriction affects growth and body composition and hepatic gene expression for nitrogen metabolism in growing rats. J Anim Physiol Anim Nutr 98:149–157

    Article  CAS  Google Scholar 

  • Křížová L, Třináctý J, Svobodová J, Richter M, Černý V, Jarošová A (2010) Effect of supplemental rumenprotected lysine, methionine or both added to diet of lactating dairy cows on milk fatty acids profile. Acta Univ Agric Silvic Mendelianae Brun 58:87–94

    Article  Google Scholar 

  • Kunio T (1998) Central mechanism of umami taste perception and effect of dietary protein on the preference for amino acids and sodium chloride in rats. Food Rev Int 14:273–308

    Article  Google Scholar 

  • Labadie J, Dunn WA, Aronson NN (1976) Hepatic synthesis of carnitine from protein-bound trimethyl-lysine. Lysosomal digestion of methyl-lysine-labelled asialo-fetuin. Biochem J 160:85–95

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lapierre H, Doepel L, Milne E, Lobley GE (2009) Responses in mammary and splanchnic metabolism to altered lysine supply in dairy cows. Animal 3:360–371

    Article  CAS  PubMed  Google Scholar 

  • Luigi S, Alfonso G, Domenico C, Domenico C, Luisa BM (2014) Where does Nε-trimethyllysine for the carnitine biosynthesis in mammals come from? PLoS ONE 9:e84589

    Article  CAS  Google Scholar 

  • Madeira MS, Costa P, Alfaia CM, Lopes PA, Bessa RJB, Lemos JPC, Prates JAM (2013) The increased intramuscular fat promoted by dietary lysine restriction in lean but not in fatty pig genotypes improves pork sensory attributes. J Anim Sci 91:3177–3187

    Article  CAS  PubMed  Google Scholar 

  • Markovitz PJ, Chuang DT, Cox RP (1984) Familial hyperlysinemias. Purification and characterization of the bifunctional aminoadipic semialdehyde synthase with lysine-ketoglutarate reductase and saccharopine dehydrogenase activities. J Biol Chem 259:11643–11646

    CAS  PubMed  Google Scholar 

  • National Research Council (2001) Nutrient requirements of dairy cattle. National Academy Press, Washington

    Google Scholar 

  • Ogunade I, Schweickart H, Mccoun M, Cannon K, Mcmanus C (2019) Integrating 16S rRNA sequencing and LC–MS-based metabolomics to evaluate the effects of live yeast on rumen function in beef cattle. Animals 9:28

    Article  Google Scholar 

  • Pink DBS, Gatrell SK, Elango R, Turchinsky J, Kiess AS, Blemings KP, Dixon WT, Ball RO (2011) Lysine α-ketoglutarate reductase, but not saccharopine dehydrogenase, is subject to substrate inhibition in pig liver. Nutrit Res 31:544–554

    Article  CAS  Google Scholar 

  • Pon CK, Lane JR, Sloan EK, Halls ML (2016) The β2-adrenoceptor activates a positive cAMP-calcium feedforward loop to drive breast cancer cell invasion. The FASEB J 30:1144–1154

    Article  CAS  PubMed  Google Scholar 

  • Rodríguez-Sánchez JA, Sanz MA, Blanco M, Serrano MP, Joy M, Latorre MA (2011) The influence of dietary lysine restriction during the finishing period on growth performance and carcass, meat, and fat characteristics of barrows and gilts intended for dry-cured ham production. J Anim Sci 89:3651–3662

    Article  PubMed  CAS  Google Scholar 

  • Schaff CT, Rohrbeck D, Steinhoff-Wagner J, Kanitz E, Sauerwein H, Bruckmaier RM, Hammon HM (2014) Effects of colostrum versus formula feeding on hepatic glucocorticoid and α(1)- and β(2)-adrenergic receptors in neonatal calves and their effect on glucose and lipid metabolism. J Dairy Sci 97:6344–6357

    Article  CAS  PubMed  Google Scholar 

  • Schneider JD, Tokach MD, Dritz SS, Nelssen JL, DeRouchey JM, Goodband RD (2010) Determining the effect of lysine:calorie ratio on growth performance of ten- to twenty-kilogram of body weight nursery pigs of two different genotypes. J Anim Sci 88:137–146

    Article  CAS  PubMed  Google Scholar 

  • Schwab CG, Broderick GA (2017) A 100-Year Review: protein and amino acid nutrition in dairy cows. J Dairy Sci 100:10094–10112

    Article  CAS  PubMed  Google Scholar 

  • Sekhar V, Pollicino T, Diaz G, Engle RE, Alayli F, Melis M, Kabat J, Tice A, Pomerenke A, Altan-Bonnet N, Zamboni F, Lusso P, Emerson SU, Farci P (2018) Infection with hepatitis C virus depends on TACSTD2, a regulator of claudin-1 and occludin highly downregulated in hepatocellular carcinoma. PloS Pathog 14:e1006916

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Servillo L, Giovane A, Diaz G, Cautela D, Cartaldo D, Balestrieri ML (2014) Where does N(ε)-trimethyllysine for the carnitine biosynthesis in mammals come from? PLoS ONE 9:e84589

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Smriga M, Murakami H, Mori M, Torii K (2000) Effects of l-lysine deficient diet on the hypothalamic interstitial norepinephrine and diet-induced thermogenesis in rats in vivo. BioFactors 12:137–142

    Article  CAS  PubMed  Google Scholar 

  • Soest PJ, Van RJB, Lewis BA (1991) Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci 74:3583–3597

    Article  PubMed  Google Scholar 

  • Swanepoel N, Robinson PH, Erasmus LJ (2010) Amino acid needs of lactating dairy cows: Impact of feeding lysine in a ruminally protected form on productivity of lactating dairy cows. Anim Feed Sci Technol 157:79–94

    Article  CAS  Google Scholar 

  • Takagi M, Yonezawa T, Haga S, Shingu H, Kobayashi Y, Takahashi T, Ohtani Y, Obara Y, Katoh K (2008) Changes of activity and mRNA expression of urea cycle enzymes in the liver of developing Holstein calves. J Anim Sci 86:1526–1532

    Article  CAS  PubMed  Google Scholar 

  • Tchagang AB, Tewfik AH, Derycke MS, Skubitz KM, Skubitz AP (2008) Early detection of ovarian cancer using group biomarkers. Mol Cancer Ther 7:27–37

    Article  CAS  PubMed  Google Scholar 

  • Tsurugizawa T, Uematsu A, Uneyama H, Torii K (2013) Reversible brain response to an intragastric load of l-lysine under l-lysine depletion in conscious rats. Br J Nutr 109:1323–1329

    Article  CAS  PubMed  Google Scholar 

  • Vels L, Røntved CM, Bjerring M, Ingvartsen KL (2009) Cytokine and acute phase protein gene expression in repeated liver biopsies of dairy cows with a lipopolysaccharide-induced mastitis. J Dairy Sci 92:922–934

    Article  CAS  PubMed  Google Scholar 

  • Wan PJ, Yuan SY, Tang YH, Li KL, Yang L, Fu Q, Li GQ (2015) Pathways of amino acid degradation in nilaparvata lugens (Stål) with special reference to lysine-ketoglutarate reductase/saccharopine dehydrogenase (LKR/SDH). PLoS ONE 10:e0127789

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Wang Z, Gerstein M, Snyder M (2009) RNA-Seq: a revolutionary tool for transcriptomics. Nat Rev Genet 10:57–63

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang J, Diao Q, Tu Y, Zhang N, Xu X (2012) The limiting sequence and proper ratio of lysine, methionine and threonine for calves fed milk replacers containing soy protein. Asian-Australas J Anim Sci 25:224–233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Williams JE, Newell SA, Hess BW, Scholljegerdes E (1999) Influence of rumen-protected methionine and lysine on growing cattle fed forage and corn based diets. J Prod Agric 12:696–701

    Article  Google Scholar 

  • Wrann CD, Eguchi J, Bozec A, Xu Z, Mikkelsen T, Gimble J, Nave H, Wagner EF, Ong SE, Rosen ED (2012) FOSL2 promotes leptin gene expression in human and mouse adipocytes. J Clin Invest 122:1010–1021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xue F, Zhou Z, Ren L, Meng Q (2011) Influence of rumen-protected lysine supplementation on growth performance and plasma amino acid concentrations in growing cattle offered the maize stalk silage/maize grain-based diet. Anim Feed Sci Technol 169:61–67

    Article  CAS  Google Scholar 

  • Zinn RA, Shen Y (1998) An evaluation of ruminally degradable intake protein and metabolizable amino acid requirements of feedlot calves. J Anim Sci 76:1280–1289

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported, in part, by the Earmarked Fund for Beijing Dairy Industry Innovation Consortium of Agriculture Research System (BAIC06-2019), Chinese Academy of Agricultural Science and Technology Innovation Project (CAAS-XTCX-2016011-01), Fundamental Research Funds for Central Non-profit Scientific Institution (Y2019CG08), The Agricultural Science and Technology Innovation Program (CAAS-ASTIP-2017-FRI-04), and Science and Technology Open Cooperation Project of Henan Province (182106000035). The authors thank Yijia Zhang (College of Veterinary Medicine, China Agricultural University) for her technical assistance in the study.

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  1. Yanliang Bi and Fanlin Kong contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Feed Biotechnology of the Ministry of Agriculture, Sino-US Joint Lab On Nutrition and Metabolism of Ruminants, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Fanlin Kong, Yanliang Bi, Kai Cui, Yuan Li, Qiyu Diao & Yan Tu

  2. Department of Grassland Science, China Agricultural University, Beijing, 100081, China

    Bing Wang

  3. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450002, China

    Tong Fu

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  1. Fanlin Kong
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Correspondence to Yan Tu.

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The experimental procedures were approved by the Animal Ethics Committee of the Chinese Academy of Agricultural sciences (committee permit number: FRI-CAAS20180110), and humane animal care and handing procedures were followed throughout the experiment.

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Kong, F., Bi, Y., Wang, B. et al. Integrating RNA-sequencing and untargeted LC–MS metabolomics to evaluate the effect of lysine deficiency on hepatic functions in Holstein calves. Amino Acids 52, 781–792 (2020). https://doi.org/10.1007/s00726-020-02852-1

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