Abstract
The genotoxic effects of a mixture of 13 volatile organic compounds (VOCs) on total DNA profiles and the expression of heat shock proteins (HSPs) HSP26, HSP60, HSP70, and HSP83 in fruit fly tissues were examined. Drosophila melanogaster Oregon R+, reared under controlled conditions on artificial diets, was treated with 13 VOCs commonly found in water at concentrations of 10, 20, 50, and 75 ppb for 1 and 5 days. Band changes were clearly detected in random amplified polymorphic DNA assay, especially at the 50- and 75-ppb exposure levels, for both treatment periods. In addition, there were clear differences in the band profiles of the treated and untreated flies with changes in the band intensity and the loss/appearance of bands. Although the genomic template stability (GTS) exhibited irregular changes at the first day, significant decreases in GTS were observed after 5 days of VOC application. The lowest GTS value (27.77 ± 1.96 %) was detected at the 75-ppb level after 5 days of the treatment. Quantitative real-time polymerase chain reaction analysis showed a significant increase in the relative expression of HSP26 and HSP60 after 1 and 5 days of the treatment, respectively. The expression of HSP70 increased significantly at all treatment concentrations and times. However, the greatest increase in expression level of HSP70 (4.2-fold) occurred at 20 ppb after 5 days of the treatment. HSP83 was the least affected by exposure to the VOCs. We conclude that trace levels of a mixture of VOCs can exert genotoxic effects on both total DNA and HSP levels in Drosophila.
Similar content being viewed by others
Explore related subjects
Discover the latest articles, news and stories from top researchers in related subjects.References
Ait-Aissa S, Porcher JM, Arrigo AP, Lambre C (2000) Activation of the hsp70 promoter by environmental inorganic and organic chemicals: relationships with cytotoxicity and lipophilicity. Toxicology 145:147–157
Andersen HR, Nielsen JB, Grandjean P (2000) Toxicologic evidence of developmental neurotoxicity of environmental chemicals. Toxicology 144:121–127
Atienzar FA, Conradi M, Evenden AJ, Jha AN, Depledge MH (1999) Qualitative assessment of genotoxicity using random amplified polymorphic DNA, comparison of genomic template stability with key fitness parameters in Daphnia magna exposed to benzo[a]pyrene. Environ Toxicol Chem 18:2275–2282
Atienzar FA, Cordi B, Donkin ME, Evenden AJ, Jha AN, Depledge MH (2000) Comparison of ultraviolet-induced genotoxicity detected by random amplified polymorphic DNA with chlorophyll fluorescence and growth in a marine macroalgae, Palmaria palmate. Aquat Toxicol 50:1–12
Atienzar FA, Billinghurst Z, Depledge MH (2002) 4-n-Nonylphenol and 17-in developing barnacle larvae. Environ Pollut 120:735–738
Baydas G, Reiter RJ, Nedzvetskii VS, Yasar A, Tuzcu M, Ozveren F et al (2003) Melatonin protects the central nervous system of rats against toluene containing thinner intoxication by reducing reactive gliosis. Toxicol Lett 137:169–174
Castaño C, Becerril P, Garcia H, Acevevedo H (2002) Detection of genotoxicity by RAPDs on fish cells after acute and chronic exposure to B[a]P. In: Proceedings of the 12th SETAC Europe Annual Meeting 233
Cenkci S, Yildiz M, Cigerci IH, Bozdag A, Terzi ESA (2010) Evaluation of 2,4-D and Dicamba genotoxicity in bean seedlings using comet and RAPD assays. Ecotoxicol Environ Saf 73(7):1558–1564
Ceyhun SB et al (2010) Deltamethrin attenuates antioxidant defense system and induces the expression of heat shock protein 70 in rainbow trout. Comp Biochem Physiol A Mol Integr Physiol 152(2):215–223
Conte C, Mutti I, Puglisi P, Ferrarini A, Regina G, Maestri E (1998) DNA fingerprinting analysis by a PCR based method for monitoring the genotoxic effects of heavy metals pollution. Chemosphere 37:2739–2749
Dauneau P, Perez-Martinez G (1997) Fractional factorial design and multiple linear regression to optimise extraction of volatiles from Lactobacillus plantarum bacterial suspension using purge and trap. J Chromatogr A 775(1):225–230
Desai NS, Agarwal AA, Uplap SS (2010) HSP: evolved and conserved proteins, structure and sequence studies. Int J Bioinform Res 2(2)
Doganlar ZB (2012) Physiological and genetic responses to pesticide mixture treatment of Veronica beccabunga. Water Air Soil Pollut 223(9):6201–6212
Doganlar ZB, Doganlar O, Tabakçıoğlu K (2014) Genotoxic effects of heavy metal mixture in Drosophila melanogaster: expressions of heat shock proteins, RAPD profiles and mitochondrial DNA sequence. Water Air Soil Pollut 225(9):1–14
Dreiem A, Myhre O, Fonnum F (2002) Relationship between lipophilicity of C6-10 hydrocarbon solvents and their ROS-inducing potency in rat cerebellar granule cells. Neurotoxicology 23:701–709
Dreiem A, Ring A, Fonnum F (2005) Organic solvent-induced cell death in rat cerebellar granule cells: structure dependence of c10 hydrocarbons and relationship to reactive oxygen species formation. Neurotoxicology 26:321–330
Duffy LK, Scofield E, Rodgers T, Patton M, Bowyer RT (1999) Comparative baseline levels of mercury, Hsp 70 and Hsp 60 in subsistence fish from the Yukon-Kuskokwim delta region of Alaska. Comp Biochem Physiol C Pharmacol Toxicol 124(2):181–186
Dunemann L, Hajimiragha H (1993) Development of a screening method for the determination of volatile organic compounds in body fluids and environmental samples using purge and trap gas chromatography-mass spectrometry. Anal Chim Acta 283(1):199–206
Eckwert H, Alberti G, Kohler HR (1997) The induction of stress proteins (hsp) in Oniscus asellus (Isopoda) as a molecular marker of multiple heavy metal exposure: I. Principles and toxicological assessment. Ecotoxicology 6(5):249–262
Gaivao I, Sierra LM, Comendator MA (1999) The w/w1 SMART assay of Drosophila melanogaster detects the genotoxic effects of reactive oxygen species inducing compounds. Mutat Res 440:139–145
Gao Q, Song L, Ni D, Wu L, Zhang H, Chang Y (2007) cDNA cloning and mRNA expression of heat shock protein 90 gene in the haemocytes of Zhikong scallop Chlamys farreri. Comp Biochem Physiol B Biochem Mol Biol 147(4):704–715
Gehrmann M, Radons J, Molls M, Multhoff G (2008) The therapeutic implications of clinically applied modifiers of heat shock protein 70 (Hsp70) expression by tumor cells. Cell Stress Chaperones 13(1):1–10
Gérin M, Siemiatychi J, Désy M, Krewski D (1998) Associations between several sites of cancer and occupational exposure to benzene, toluene, xylene, and styrene: results of a case-control study in Montreal. Am J Ind Med 34:144–156
Gupta SC, Siddique HR, Mathur N, Mishra RK, Mitra K, Saxena DK et al (2007) Adverse effect of organophosphate compounds dichlorvos and chlorpyrifos in the reproductive tissues of transgenic Drosophila melanogaster: 70 kDa heat shock protein as a marker of cellular damage. Toxicology 238(1):1–14
Gupta SC, Sharma A, Mishra M, Mishra RK, Chowdhuri DK (2010) Heat shock proteins in toxicology: how close and how far? Life Sci 86(11):377–384
Hunter ES, Dix D (1996) Antisense oligonucleotides against Hsp70-1 and Hsp70-3 increase mouse embryonic sensitivity to arsenite induced dysmorphogenesis in vitro. Teratology 53:86
International Agency for Research on Cancer (1982) IARC monographs on the evaluation of the carcinogenic risk of chemicals to humans. In: Some industrial chemical dyestuffs. vol. 29, International Agency for Research on Cancer, Lyon France, pp 93–148
Iwama GK, Afonso LO, Todgham A, Ackerman P, Nakano K (2004) Are hsps suitable for indicating stressed states in fish? J Exp Biol 207(1):15–19
Jimi S, Uchiyama M, Takaki A (2004) Mechanisms of cell death induced by cadmium and arsenic. Ann N Y Acad Sci 101:325–331
Jones C, Kortenkamp A (2000) RAPD library fingerprinting of bacterial and human DNA: applications in mutation detection. Teratog Carcinog Mutagen 20:49–63
Kim JH, Moon JY, Park EY, Lee KH, Hong YC (2011) Changes in oxidative stress biomarker and gene expression levels in workers exposed to volatile organic compounds. Ind Health 49(1):8–14
Koh K, Evans JM, Hendricks JC, Sehagal A (2006) A Drosophila model for age-associated changes in sleep:wake cycles. Proc Natl Acad Sci U S A 103:13843–13847
Lee SB, Choi J (2006) Multilevel evaluation of nonylphenol toxicity in fourth-instar larvae of Chironomus riparius (Diptera, Chironomidae). Environ Toxicol Chem 25(11):3006–3014
Lee KJ, Terada K, Oyadomari S, Inomata Y, Mori M, Gotoh T (2004) Induction of molecular chaperones in carbon tetrachloride-treated rat liver: implications in protection against liver damage. Cell Stress Chaperones 9(1):58–68
Lindquist S, Craig EA (1988) The heat-shock proteins. Annu Rev Genet 22(1):631–677
Liu W, Li PJ, Qi XM, Zhou QX, Zheng L, Sun TH (2005) DNA changes in barley (Hordeum vulgare) seedlings induced by cadmium pollution using RAPD analysis. Chemosphere 61:158–167
Lundberg I, Milatou-Smith R (1998) Mortality and cancer incidence among Swedish paint industry workers with long-term exposure to organic solvents. Scand J Work Environ Health 24:270–275
Maroni M, Seifert B, Lindvall T (1995) Indoor air quality—a comprehensive reference book. Elsevier, Amsterdam
Migliore L, Fontana I, Colognato R, Coppede F, Siciliano G, Murri L (2005) Searching for the role and the most suitable biomarkers of oxidative stress in Alzheimer’s disease and in other neurodegenerative diseases. Neurobiol Aging 26(5):587–595
Mølhave L (1991) Volatile organic compounds, indoor air quality and health. Indoor Air 1(4):357–376
Morey M, Serras F, Corominas M (2003) Halving the selenophosphate synthetase gene dose confers hypersensitivity to oxidative stress in Drosophila melanogaster. FEBS Lett 534:111–114
Myhre O, Fonnum F (2001) The effect of aliphatic, naphthenic, and aromatic hydrocarbons on production of reactive oxygen species and reactive nitrogen species in rat brain synaptosome fraction: the involvement of calcium, nitric oxide synthase, mitochondria, and phospholipase. Biochem Pharmacol 62:119–128
Norbäck D, Björnsson E, Janson C, Widström J, Boman G (1995) Asthmatic symptoms and volatile organic compounds, formaldehyde, and carbon dioxide in dwellings. Occcup Environ Med 52:388–395
Ozturk F, Duman F, Leblebici Z, Temizgul R (2010) Arsenic accumulation and biological responses of watercress (Nasturtium officinale R. Br.) exposed to arsenite. Environ Exp Bot 69(2):167–174
Piano A, Valbonesi P, Fabbri E (2004) Expression of cytoprotective proteins, heat shock protein 70 and metallothioneins, in tissues of Ostrea edulis exposed to heat and heavy metals. Cell Stress Chaperones 9(2):134
Posgai R, Cipolla-McCulloch CB, Murphy KR, Hussain SM, Rowe JJ, Nielsen MG (2011) Differential toxicity of silver and titanium dioxide nanoparticles on Drosophila melanogaster development, reproductive effort, and viability: size, coatings and antioxidants matter. Chemosphere 85:34–42
Pyza E, Mak P, Kramarz P, Laskowski R (1997) Heat shock proteins (HSP70) as biomarkers in ecotoxicological studies. Ecotoxicol Environ Saf 38(3):244–251
Qian Z, Liu X, Wang L, Wang X, Li Y, Xiang J et al (2012) Gene expression profiles of four heat shock proteins in response to different acute stresses in shrimp, Litopenaeus vannamei. Comp Biochem Physiol C Pharmacol Toxicol 156(3):211–220
Reiter LT, Potocki L, Chien S, Gribskov M, Bier E (2001) A systematic analysis of human disease-associated gene sequences in Drosophila melanogaster. Genome Res 11:1114–1125
Republic of Turkey Ministry of Health (2005) Regulation on water intended for human consumption. 23 March 2013/28580
Roh JY, Lee J, Choi J (2006) Assessment of stress-related gene expression in the heavy metal-exposed nematode Caenorhabditis elegans: A potential biomarker for metal-induced toxicity monitoring and environmental risk assessment. Environ Toxicol Chem 25(11):2946–2956
Rossner JR, Binkova B, Sram RJ (2003) Heat shock proteins hsp32 and hsp70 as biomarkers of an early response? In vitro induction of heat shock proteins after exposure of cell culture to carcinogenic compounds and their real mixtures. Mutat Res 542(1–2):105–116
Ryan JA, Hightower LE (1996) Stress proteins as molecular biomarkers for environmental toxicology. In: Friege U, Morimoto RI, Yahara I, Polla B (eds) Stress-inducible cellular responses. Birkhäuser, Basel, pp 411–424
Said A, Ferencz K, Nemcsók AJ, Hermesz E (2010) Expressions of heat shock and metallothionein genes in the heart of common carp (Cyprinus carpio): effects of temperature shock and heavy metal exposure. Acta Biol Hung 61(1):10–23
Siddique HR, Chowdhuri DK, Saxena DK, Dhawan A (2005) Validation of Drosophila melanogaster as an in vivo model for genotoxicity assessment using modified alkaline Comet assay. Mutagenesis 20(4):285–290
Silins I, Hogberg J (2011) Combined toxic exposures and human health: biomarkers of exposure and effect. Int J Environ Res Publ Health 8(3):629–647
Singh MP, Reddy MM, Mathur N, Saxena DK, Chowdhuri DK (2009) Induction of hsp70, hsp60, hsp83 and hsp26 and oxidative stress markers in benzene, toluene and xylene exposed Drosophila melanogaster: role of ROS generation. Toxicol Appl Pharmacol 235(2):226–243
Singh MP, Ravi Ram K, Mishra M, Shrivastava M, Saxena DK, Chowdhuri DK (2010) Effects of co-exposure of benzene, toluene and xylene to Drosophila melanogaster: alteration in hsp70, hsp60, hsp83, hsp26, ROS generation and oxidative stress markers. Chemosphere 79(5):577–587
Singh MP, Mishra M, Sharma A, Shukla AK, Mudiam MKR, Patel DK et al (2011) Genotoxicity and apoptosis in Drosophila melanogaster exposed to benzene, toluene and xylene: attenuation by quercetin and curcumin. Toxicol Appl Pharmacol 253(1):14–30
Snyder MJ, Girvetz E, Mulder EP (2001) Induction of marine mollusc stress proteins by chemical or physical stress. Arch Environ Contam Toxicol 41:22–29
Spiller HA, Krenzelok EP (1997) Epidemiology of inhalant abuse reported to two regional poison centers. J Toxicol Clin Toxicol 35:167–173
Stone V, Wilson MR, Lightbody J, Donaldson K (2003) Investigating the potential for interaction between the components of PM10. Environ Health Prev Med 7(6):246–253
Swaileh KM, Hussein RA, Ezzughayyar A (2008) Evaluating wastewater-induced plant genotoxicity using randomly amplified polymorphic DNA. Environ Toxicol 23(1):117–122
Tsuda M, Ootaka R, Ohkura C, Kishita Y, Seong KH, Matsuo T et al (2010) Loss of Trx-2 enhances oxidative stress-dependent phenotypes in Drosophila. FEBS Lett 584(15):3398–3401
United States Environmental Protection Agency (2009) National primary drinking water regulations EPA 816-F-09-0004
United States Office of the Federal Register (1995) Code of federal regulations, vol 21. US Government Printing Office, Washington, DC, pp 170–199
Vijayan MM, Pereira C, Forsyth RB, Kennedy CJ, Iwama GK (1997) Handling stress does not affect the expression of hepatic heat shock protein 70 and conjugation enzymes in rainbow trout treated with beta-naphthoflavone. Life Sci 61(2):117–127
Waisberg M et al (2003) Molecular and cellular mechanisms of cadmium carcinogenesis. Toxicology 192(2):95–117
Warrick JM, Chan HY, Gray-Board GL, Chai Y, Paulson HL, Bonini NM (1999) Suppression of polyglutamine mediated neurodegeneration in Drosophila by the molecular chaperone HSP70. Nat Genet 23:425–428
Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18(22):6531–6535
Wolf MJ, Amrein H, Izatt JA, Choma MA, Reedy MC, Rockman HA (2006) Drosophila as a model for the identification of genes causing adult human heart disease. PNAS 103:1394–1399
Zarate J, Bradley TM (2003) Heat shock proteins are not sensitive indicators of hatchery stress in salmon. Aquaculture 223(1):175–187
Acknowledgments
The author is grateful to Dr. Yener Yoruk (Rector of Trakya University, Edirne/Turkey) and the Technology Research and Application Centre, which is funded by T. R. State Planning Organization (Project No. 2011K120390) for providing laboratory equipment.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Doganlar, O., Doganlar, Z.B. Effects of a Mixture of Volatile Organic Compounds on Total DNA and Gene Expression of Heat Shock Proteins in Drosophila melanogaster . Arch Environ Contam Toxicol 68, 395–404 (2015). https://doi.org/10.1007/s00244-014-0089-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00244-014-0089-4