Skip to main content

Dietary l-arginine accelerates pupation and promotes high protein levels but induces oxidative stress and reduces fecundity and life span in Drosophila melanogaster

Abstract

l-Arginine, a precursor of many amino acids and of nitric oxide, plays multiple important roles in nutrient metabolism and regulation of physiological functions. In this study, the effects of l-arginine-enriched diets on selected physiological responses and metabolic processes were assessed in Drosophila melanogaster. Dietary l-arginine at concentrations 5–20 mM accelerated larval development and increased body mass, and total protein concentrations in third instar larvae, but did not affect these parameters when diets contained 100 mM arginine. Young (2 days old) adult flies of both sexes reared on food supplemented with 20 and 100 mM l-arginine possessed higher total protein concentrations and lower glucose and triacylglycerol concentrations than controls. Additionally, flies fed 20 mM l-arginine had higher proline and uric acid concentrations. l-Arginine concentration in the diet also affected oxidative stress intensity in adult flies. Food with 20 mM l-arginine promoted lower protein thiol concentrations and higher catalase activity in flies of both sexes and higher concentrations of low molecular mass thiols in males. When flies were fed on a diet with 100 mM l-arginine, lower catalase activities and concentrations of protein thiols were found in both sexes as well as lower low molecular mass thiols in females. l-Arginine-fed males demonstrated higher climbing activity, whereas females showed higher cold tolerance and lower fecundity, compared with controls. Food containing 20 mM l-arginine shortened life span in both males and females. The results suggest that dietary l-arginine shows certain beneficial effects at the larval stage and in young adults. However, the long-term consumption of l-arginine-enriched food had unfavorable effects on D. melanogaster due to decreasing fecundity and life span.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Abbreviations

ABTS:

2,2-Azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

GDH:

Glutamate dehydrogenase

FSA:

Food supplemented with l-arginine

H-SH:

High molecular mass thiol groups

KPi:

Potassium phosphate buffer

L-SH:

Low molecular mass thiol groups

ROS:

Reactive oxygen species

TAG:

Triacylglycerols

References

  • Aebi H (1984) Catalase in vitro. Meth Enzymol 105:121–126

    Article  CAS  PubMed  Google Scholar 

  • Andersen LH, Kristensen TN, Loeschcke V, Toft S, Mayntz D (2010) Protein and carbohydrate composition of larval food affects tolerance to thermal stress and desiccation in adult Drosophila melanogaster. J Insect Physiol 56:336–340. doi:10.1016/j.jinsphys.2009.11.006

    Article  CAS  PubMed  Google Scholar 

  • Bauchart-Thevret C, Cui L, Wu G, Burrin DG (2010) Arginine-induced stimulation of protein synthesis and survival in IPEC-J2 cells is mediated by mTOR but not nitric oxide. Am J Physiol Endocrinol Metab 299:E899–E909. doi:10.1152/ajpendo.00068.2010

    Article  CAS  PubMed  Google Scholar 

  • Bayliak MM, Shmihel HV, Lylyk MP, Vytvytska OM, Storey JM, Storey KB, Lushchak VI (2015) Alpha-ketoglutarate attenuates toxic effects of sodium nitroprusside and hydrogen peroxide in Drosophila melanogaster. Environ Toxicol Pharmacol 40:650–659. doi:10.1016/j.etap.2015.08.016

    Article  CAS  PubMed  Google Scholar 

  • Bayliak MM, Lylyk MP, Shmihel HV, Sorochynska OM, Manyukh OV, Pierzynowski SG, Lushchak VI (2016) Dietary alpha-ketoglutarate increases cold tolerance in Drosophila melanogaster and enhances protein pool and antioxidant defense in sex-specific manner. J Therm Biol 60:1–11. doi:10.1016/j.jtherbio.2016.06.001

    Article  CAS  PubMed  Google Scholar 

  • Bergman I, Loxley R (1970) New spectrophotometric method for the determination of proline in tissue hydrolyzates. Anal Chem 42:702–706

    Article  CAS  PubMed  Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Cáceres L, Necakov AS, Schwartz C, Kimber S, Roberts IJ, Krause HM (2011) Nitric oxide coordinates metabolism, growth, and development via the nuclear receptor E75. Genes Dev 25:1476–1485. doi:10.1101/gad.2064111

    Article  PubMed  PubMed Central  Google Scholar 

  • Cao W, Xiao L, Liu G, Fang T, Wu X, Jia G, Zhao H, Chen X, Wu C, Cai J, Wang J (2016) Dietary arginine and N-carbamylglutamate supplementation enhances the antioxidant statuses of the liver and plasma against oxidative stress in rats. Food Funct 7:2303–2311. doi:10.1039/c5fo01194a

    Article  CAS  PubMed  Google Scholar 

  • Chamruspollert M, Pesti GM, Bakalli RI (2002) Dietary interrelationships among arginine, methionine, and lysine in young broiler chicks. Br J Nutr 88:655–660

    Article  CAS  PubMed  Google Scholar 

  • Chapman RF, Simpson SJ, Douglas AE (2013) The insects: structure and function. Cambridge University Press. http://books.google.co.uk/books?id=NXJEi8fo7CkC

  • David RJ, Gibert P, Pla E, Petavy G, Karan D, Moreteau B (1998) Cold stress tolerance in Drosophila: analysis of chill coma recovery in D. melanogaster. J Therm Biol 23:291–299

    Article  Google Scholar 

  • Davies S (2000) Nitric oxide signaling in insects. Insect Biochem Mol Biol 30:1123–1138

    Article  CAS  PubMed  Google Scholar 

  • Delalio LJ, Dion SM, Bootes AM, Smith WA (2015) Direct effects of hypoxia and nitric oxide on ecdysone secretion by insect prothoracic glands. J Insect Physiol 76:56–66. doi:10.1016/j.jinsphys.2015.02.009

    Article  CAS  PubMed  Google Scholar 

  • Doherty D (1970) l-Glutamate dehydrogenases (yeast). Meth Enzymol 17:850–856

    Article  Google Scholar 

  • Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77

    Article  CAS  PubMed  Google Scholar 

  • Erel O (2004) A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem 37(4):277–285

    Article  CAS  PubMed  Google Scholar 

  • Gospodaryov DV, Yurkevych IS, Lushchak OV, Lushchak VI (2013) Correction: Lifespan extension and delay of age-related functional decline caused by Rhodiola rosea depends on dietary macronutrient balance. Longev Healthspan 2(1):12. doi:10.1186/2046-2395-2-12

    Article  PubMed  PubMed Central  Google Scholar 

  • Grandison RC, Piper MD, Partridge L (2009) Amino-acid imbalance explains extension of lifespan by dietary restriction in Drosophila. Nature 462(7276):1061–1064. doi:10.1038/nature08619

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Harper CJ, Hayward D, Kidd M, Wiid I, van Helden P (2010) Glutamate dehydrogenase and glutamine synthetase are regulated in response to nitrogen availability in Myocbacterium smegmatis. BMC Microbiol 10:138. doi:10.1186/1471-2180-10-138

    Article  PubMed  PubMed Central  Google Scholar 

  • Hess DT, Matsumoto A, Kim SO, Marshall HE, Stamler JS (2005) Protein S-nitrosylation: purview and parameters. Nat Rev Mol Cell Biol 6(2):150–166

    Article  CAS  PubMed  Google Scholar 

  • Hilliker AJ, Duyf B, Evans D, Phillips JP (1992) Urate-null rosy mutants of Drosophila melanogaster are hypersensitive to oxygen stress. Proc Natl Acad Sci 89(10):4343–4347

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hinton T (1956) The effects of arginine, ornithine and citrulline on the growth of Drosophila. Arch Biochem Biophys 62(1):78–85

    Article  CAS  PubMed  Google Scholar 

  • Jobgen W, Meininger CJ, Jobgen SC, Li P, Lee MJ, Smith SB, Spencer TE, Fried SK, Wu G (2009) Dietary l-arginine supplementation reduces white fat gain and enhances skeletal muscle and brown fat masses in diet-induced obese rats. J Nutr 139(2):230–237. doi:10.3945/jn.108.096362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jones MA, Grotewiel M (2011) Drosophila as a model for age-related impairment in locomotor and other behaviors. Exp Gerontol 46:320–325

    Article  PubMed  Google Scholar 

  • Kapahi P, Zid BM, Harper T, Koslover D, Sapin V, Benzer S (2004) Regulation of lifespan in Drosophila by modulation of genes in the TOR signaling pathway. Curr Biol 14:885–890

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kocna P, Frych P, Zavoral MT, Pelech T (1996) Arginase activity determination. A marker of large bowel mucosa proliferation. Eur J Clin Chem Clin Biochem 34(8):619–623

    CAS  PubMed  Google Scholar 

  • Koštál V, Korbelová J, Poupardin R, Moos M, Šimek P (2016) Arginine and proline applied as food additives stimulate high freeze tolerance in larvae of Drosophila melanogaster. J Exp Biol 219:2358–2367. doi:10.1242/jeb.142158

    Article  PubMed  Google Scholar 

  • Kowalski S, Aubin T, Martin Can JR (2004) Courtship song in Drosophila melanogaster: a differential effect on male–female locomotor activity. J Zool 82:1258–1266

    Google Scholar 

  • Kraaijeveld AR, Elrayes NP, Schuppe H, Newland PL (2011) L-arginine enhances immunity to parasitoids in Drosophila melanogaster and increases NO production in lamellocytes. Dev Comp Immunol 35:857–864. doi:10.1016/j.dci.2011.03.019

    Article  CAS  PubMed  Google Scholar 

  • Kuzin B, Roberts I, Peunova N, Enikolopov G (1996) Nitric oxide regulates cell proliferation during Drosophila development. Cell 87(4):639–649

    Article  CAS  PubMed  Google Scholar 

  • Lee YP, Takahashi T (1966) An improved colorimetric determination of amino acids with the use of ninhydrin. Anal Biochem 14(1):71–77

    Article  CAS  Google Scholar 

  • Lozinsky OV, Lushchak OV, Storey JM, Storey KB, Lushchak VI (2012) Sodium nitroprusside toxicity in Drosophila melanogaster: delayed pupation, reduced adult emergence, and induced oxidative/nitrosative stress in eclosed flies. Arch Insect Biochem Physiol 80(3):166–185. doi:10.1002/arch.21033

    Article  CAS  PubMed  Google Scholar 

  • Lozinsky OV, Lushchak OV, Kryshchuk NI, Shchypanska NY, Riabkina AH, Skarbek SV, Maksymiv IV, Storey JM, Storey KB, Lushchak VI (2013) S-nitrosoglutathione-induced toxicity in Drosophila melanogaster: Delayed pupation and induced mild oxidative/nitrosative stress in eclosed flies. Comp Biochem Physiol A Mol Integr Physiol 164(1):162–170. doi:10.1016/j.cbpa.2012.08.006

    Article  CAS  PubMed  Google Scholar 

  • Lushchak VI (2012) Glutathione homeostasis and functions: potential targets for medical interventions. J Amino Acids. doi:10.1155/2012/736837

    PubMed  PubMed Central  Google Scholar 

  • Lushchak VI (2014) Free radicals, reactive oxygen species, oxidative stress and its classification. Chem Biol Interact 224:164–175. doi:10.1016/j.cbi.2014.10.016

    Article  CAS  PubMed  Google Scholar 

  • Lushchak OV, Lushchak VI (2008) Catalase modifies yeast Saccharomyces cerevisiae response towards S-nitrosoglutathione-induced stress. Redox Rep 13(6):283–291. doi:10.1179/135100008X309037

    Article  CAS  PubMed  Google Scholar 

  • Lushchak OV, Inoue Y, Lushchak VI (2010) Regulatory protein Yap1 is involved in response of yeast Saccharomyces cerevisiae to nitrosative stress. BioChemistry 75(5):629–664

    CAS  PubMed  Google Scholar 

  • Lushchak OV, Rovenko BM, Gospodaryov DV, Lushchak VI (2011) Drosophila melanogaster larvae fed by glucose and fructose demonstrate difference in oxidative stress markers and antioxidant enzymes of adult flies. Comp Biochem Physiol A Mol Integr Physiol 160(1):27–34

    Article  CAS  PubMed  Google Scholar 

  • Lushchak OV, Gospodaryov DV, Rovenko BM, Glovyak AD, Yurkevych IS, Klyuba VP, Shcherbij MV, Lushchak VI (2012) Balance between macronutrients affects life span and functional senescence in fruit fly Drosophila melanogaster. J Gerontol A Biol Sci Med Sci 67(2):118–125. doi:10.1093/gerona/glr184

    Article  PubMed  Google Scholar 

  • MacMillan HA, Williams CM, Staples JF, Sinclair BJ (2012) Reestablishment of ion homeostasis during chill-coma recovery in the cricket Gryllus pennsylvanicus. Proc Natl Acad Sci 109(50): 20750–20755. doi:10.1073/pnas.1212788109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McKnight JR, Satterfield MC, Jobgen WS, Smith SB, Spencer TE, Meininger CJ, McNeal CJ, Wu G (2010) Beneficial effects of L-arginine on reducing obesity: potential mechanisms and important implications for human health. Amino Acids 39(2):349–357. doi:10.1007/s00726-010-0598-z

    Article  CAS  PubMed  Google Scholar 

  • Mirth CK, Shingleton AW (2014) The roles of juvenile hormone, insulin/target of rapamycin, and ecdysone signaling in regulating body size in Drosophila. Commun Integr Biol 7(5):e971568. doi:10.4161/cib.29240

    Article  PubMed Central  Google Scholar 

  • Misener SR, Chen C, Walker VK (2001) Cold tolerance and proline metabolic gene expression in Drosophila melanogaster. J Insect Physiol 47(4–5):393–400

    Article  CAS  PubMed  Google Scholar 

  • Morozkina NV, Gaston B (2012) S-Nitrosylation signaling regulates cellular protein interactions. Biochim Biophys Acta 1820(6):722–729. doi:10.1016/j.bbagen.2011.06.017

    Article  Google Scholar 

  • Nappi AJ, Vass E, Frey F, Carton Y (2000) Nitric oxide involvement in Drosophila immunity. Nitric Oxide 4(4):423–430

    Article  CAS  PubMed  Google Scholar 

  • Naton JL (2015) Insect Physiology and Biochemistry, 3th Ed. CRC Press, Boca Raton

    Google Scholar 

  • Privat C, Lantoine F, Bedioui F, Millanvoye van Brussel E, Devynck J, Devynck MA (1997) Nitric oxide production by endothelial cells: comparison of three methods of quantification. Life Sci 61:1195–1202

    Article  Google Scholar 

  • Regulski M, Stasiv Y, Tully T, Enikolopov G (2004) Essential function of nitric oxide synthase in Drosophila. Curr Biol 4:R881–R882

    Article  Google Scholar 

  • Rovenko BM, Perkhulyn NV, Lushchak OV, Storey JM, Storey KB, Lushchak VI (2014) Molybdate partly mimics insulin-promoted metabolic effects in Drosophila melanogaster. Comp Biochem Physiol C Toxicol Pharmacol 165:76–82. doi:10.1016/j.cbpc.2014.06.002

    Article  CAS  PubMed  Google Scholar 

  • Rovenko BM, Kubrak OI, Gospodaryov DV, Perkhulyn NV, Yurkevych IS, Sanz A, Lushchak OV, Lushchak VI (2015) High sucrose consumption promotes obesity whereas its low consumption induces oxidative stress in Drosophila melanogaster. J Insect Physiol 79:42–54. doi:10.1016/j.jinsphys.2015.05.007

    Article  CAS  PubMed  Google Scholar 

  • Sacktor B, Childress CC (1967) Metabolism of proline in insect flight muscle and its significance in stimulating the oxidation of pyruvate. Arch Biochem Biophys 120:583–588

    Article  CAS  Google Scholar 

  • Samson ML (2000) Drosophila arginase is produced from a nonvital gene that contains the elav locus within its third intron. J Biol Chem 275:31107–31114

    Article  CAS  PubMed  Google Scholar 

  • Szabó C, Southan GJ, Thiemermann C, Vane JR (1994) The mechanism of the inhibitory effect of polyamines on the induction of nitric oxide synthase: role of aldehyde metabolites. Br J Pharmacol 113:757–766

    Article  PubMed  PubMed Central  Google Scholar 

  • Trinh I, Boulianne GL (2013) Modeling obesity and its associated disorders in Drosophila. Physiology (Bethesda) 28(2):117–124. doi:10.1152/physiol.00025.2012

    CAS  Google Scholar 

  • Vasylkovska R, Petriv N, Semchyshyn H (2015) Carbon sources for yeast growth as a precondition of hydrogen peroxide induced hormetic phenotype. Int J Microbiol. doi:10.1155/2015/697813

    PubMed  PubMed Central  Google Scholar 

  • Wildemann B, Bicker G (1999) Nitric oxide and cyclic GMP induce vesicle release at Drosophila neuromuscular junction. J Neurobiol 39:337–346

    Article  CAS  PubMed  Google Scholar 

  • Wink DA, Hines HB, Cheng RY, Switzer CH, Flores-Santana W, Vitek MP, Ridnour LA, Colton CA (2011) Nitric oxide and redox mechanisms in the immune response. J Leukoc Biol 89:873–891. doi:10.1189/jlb.1010550

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu G, Morris SM (1998) Arginine metabolism: nitric oxide and beyond. Biochem J 336:1–17

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu G, Bazer FW, Davis TA, Kim SW, Li P, Marc Rhoads J, Carey Satterfield M, Smith SB, Spencer TE, Yin Y (2009) Arginine metabolism and nutrition in growth, health and disease. Amino Acids 37(1):153–168. doi:10.1007/s00726-008-0210-y

    Article  CAS  PubMed  Google Scholar 

  • Wu G, Bazer FW, Satterfield MC, Li X, Wang X, Johnson GA, Burghardt RC, Dai Z, Wang J, Wu Z (2013) Impacts of arginine nutrition on embryonic and fetal development in mammals. Amino Acids 45:241–256. doi:10.1007/s00726-013-1515-z

    Article  CAS  PubMed  Google Scholar 

  • Yamanaka N, O’Connor MB (2011) Nitric oxide directly regulates gene expression during Drosophila development: need some gas to drive into metamorphosis? Genes Dev 25:1459–1463

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yao K, Yin YL, Chu W, Liu Z, Deng D, Li T, Huang R, Zhang J, Tan B, Wang W, Wu G (2008) Dietary arginine supplementation increases mTOR signaling activity in skeletal muscle of neonatal pigs. J Nutr 138:867–872

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Bloomington Stock Center (Indiana University, USA) for providing the D. melanogaster strain, and students V. Ivasyshyn, R. Knyhynytska, and O. Hrynkiv for technical assistance. Two anonymous reviewers are acknowledged for their highly professional and helpful recommendations for improving the paper.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Maria M. Bayliak or Volodymyr I. Lushchak.

Ethics declarations

Conflict of interest

The authors have declared that there is no conflict of interest.

Additional information

Communicated by H.V. Carey.

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bayliak, M.M., Lylyk, M.P., Maniukh, O.V. et al. Dietary l-arginine accelerates pupation and promotes high protein levels but induces oxidative stress and reduces fecundity and life span in Drosophila melanogaster . J Comp Physiol B 188, 37–55 (2018). https://doi.org/10.1007/s00360-017-1113-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00360-017-1113-6

Keywords