The impact of mutations in four essential genes involved in dopamine (DA) synthesis and transport on longevity, motor behavior, and resistance to oxidative stress was monitored in Drosophila melanogaster. The fly lines used for this study were: (i) a loss of function mutation in Catecholamines up (Catsup(26)), which is a negative regulator of the rate limiting enzyme for DA synthesis, (ii) a mutant for the gene pale (ple(2)) that encodes for the rate limiting enzyme tyrosine hydroxylase (TH), (iii) a mutant for the gene Punch (Pu(Z22)) that encodes guanosine triphosphate cyclohydrolase, required for TH activity, and (iv) a mutant in the vesicular monoamine transporter (VMAT(Δ14)), which is required for packaging of DA as vesicles inside DA neurons. Median lifespans of ple(2), Pu(Z22) and VMAT(Δ14) mutants were significantly decreased compared to Catsup(26) and wild type controls that did not significantly differ between each other. Catsup(26) flies survived longer when exposed to hydrogen peroxide (80 μM) or paraquat (10mM) compared to ple(2), Pu(Z22) or VMAT(Δ14) and controls. These flies also exhibited significantly higher negative geotaxis activity compared to ple(2), Pu(Z22), VMAT(Δ14) and controls. All mutant flies demonstrated rhythmic circadian locomotor activity in general, albeit Catsup(26) and VMAT(Δ14) flies had slightly weaker rhythms. Expression analysis of some key antioxidant genes revealed that glutathione S-transferase Omega-1 (GSTO1) expression was significantly up-regulated in all DA synthesis pathway mutants and especially in Catsup(26) and VMAT(Δ14) flies at both mRNA and protein levels. Taken together, we hypothesize that DA could directly influence GSTO1 transcription and thus play a significant role in the regulation of response to oxidative stress. Additionally, perturbations in DA synthesis do not appear to have a significant impact on circadian locomotor activity rhythms per se, but do have an influence on general locomotor activity levels.

Department of Biochemistry Molecular Biology Entomology and Plant Pathology Mississippi State University Mississippi State MS 39762 USA

Department of Biochemistry Molecular Biology Entomology and Plant Pathology Mississippi State University Mississippi State MS 39762 USA; Institute of Entomology Biology Centre Academy of Sciences and Faculty of Science University of South Bohemia Branišovská 31 370 05 České Budějovice Czech Republic

Department of Biological Sciences The University of Alabama Tuscaloosa AL 35487 USA

Department of Natural Sciences Stinson Mathematics and Science Building 3601 Stillman Blvd Stillman College Tuscaloosa AL 35043 USA

Department of Physiology and Biomedical Engineering Mayo Clinic School of Medicine Mayo Clinic 200 1st St SW Rochester MN 55905 USA

See more in PubMed

Abercrombie ED, Keefe KA, DiFrischia DS, Zigmond MJ. Differential effect of stress on in vivo dopamine release in striatum, nucleus accumbens, and medial frontal cortex. J. Neurochem. 1989;52:1655–1658. PubMed

Barron AB, Søvik E, Cornish JL. The roles of dopamine and related compounds in reward-seeking behavior across animal phyla. Front. Behav. Neurosci. 2010;4:163. eCollection. PubMed PMC

Botella JA, Bayersdorfer F, Schneuwly S. Superoxide dismutase overexpression protects dopaminergic neurons in a Drosophila model of Parkinson’s disease. Neurobiol. Dis. 2008;30:65–73. PubMed

Carbone MA, Jordan KW, Lyman RF, Harbison ST, Leips J, Morgan TJ, De Luca M, Awadalla P, Mackay TF. Phenotypic variation and natural selection at Catsup, a pleiotropic quantitative trait gene in Drosophila. Curr. Biol. 2006;16:912–919. PubMed PMC

Chaudhuri A, Bowling K, Funderburk C, Lawal H, Inamdar A, Wang Z, O'Donnell JM. Interaction of genetic and environmental factors in a Drosophila parkinsonism model. J. Neurosci. 2007;27:2457–2467. PubMed PMC

Civelli O, Bunzow JR, Grandy DK. Molecular diversity of the dopamine receptors. Annu. Rev. Pharmacol. Toxicol. 1993;33:281–307. PubMed

De Luca M, Roshina NV, Geiger-Thornsberry GL, Lyman RF, Pasyukova EG, Mackay TF. Dopa decarboxylase (Ddc) affects variation in Drosophila longevity. Nat. Genet. 2003;34:429–433. PubMed

Graham DG. Oxidative pathways for catecholamines in the genesis of neuromelanin and cytotoxic quinones. Mol. Pharmacol. 1978;14:633–643. PubMed

Greer CL, Grygoruk A, Patton DE, Ley B, Romero-Calderon R, Chang HY, Houshyar R, Bainton RJ, Diantonio A, Krantz DE. A splice variant of the Drosophila vesicular monoamine transporter contains a conserved trafficking domain and functions in the storage of dopamine, serotonin, and octopamine. J. Neurobiol. 2005;64:239–258. PubMed

Gruntenko NE, Laukhina OV, Bogomolova EV, Karpova EK, Menshanov PN, Romanova IV, Rauschenbach I.Yu. Downregulation of the dopamine D2-like receptor in corpus allatum affects juvenile hormone synthesis in Drosophila melanogaster females. J. Insect Physiol. 2012;58:348–355. PubMed

Hald A, Lotharius J. Oxidative stress and inflammation in Parkinson's disease: is there a causal link? Exp. Neurol. 2005;193:279–290. PubMed

Harbison ST, Carbone MA, Ayroles JF, Stone EA, Lyman RF, Mackay TF. Co-regulated transcriptional networks contribute to natural genetic variation in Drosophila sleep. Nat. Genet. 2009;41:371–375. PubMed PMC

Hirsh J, Riemensperger T, Coulom H, Iche M, Coupar J, Birman S. Roles of dopamine in circadian rhythmicity and extreme light sensitivity of circadian entrainment. Curr. Biol. 2010;20:209–214. PubMed PMC

Hood S, Cassidy P, Cossette MP, Weigl Y, Verwey M, Robinson B, Stewart J, Amir S. Endogenous dopamine regulates the rhythm of expression of the clock protein PER2 in the rat dorsal striatum via daily activation of D2 dopamine receptors. J. Neurosci. 2010;30:14046–14058. PubMed PMC

Hsouna A, Lawal HO, Izevbaye I, Hsu T, O'Donnell JM. Drosophila dopamine synthesis pathway genes regulate tracheal morphogenesis. Dev. Biol. 2007;308:30–43. PubMed PMC

Jakel RJ, Kern JT, Johnson DA, Johnson JA. Induction of the protective antioxidant response element pathway by 6-hydroxydopamine in vivo and in vitro. Toxicol. Sci. 2005;87:176–186. PubMed

Kim K, Kim S-H, Kim J, Kim H, Yim J. Glutathione S-transferase omega 1 activity is sufficient to suppress neurodegeneration in a Drosophila model of Parkinson disease. J. Biol. Chem. 2012;287:6628–6641. PubMed PMC

Krishnan N, Davis AJ, Giebultowicz JM. Circadian regulation of response to oxidative stress in Drosophila melanogaster. Biochem Biophys Res Commun. 2008;374:299–303. PubMed PMC

Krishnan N, Rakshit K, Chow ES, Wentzell JS, Kretzschmar. D. Giebultowicz. J.M. Loss of circadian clock accelerates aging in neurodegeneration-prone mutants. Neurobiol. Dis. 2012;45:1129–1135. PubMed PMC

Kumar S, Chen D, Sehgal A. Dopamine acts through cryptochrome to promote acute arousal in Drosophila. Genes. Dev. 2012;26:1224–1234. PubMed PMC

Kume K, Kume S, Park SK, Hirsh J, Jackson FR. Dopamine is a regulator of arousal in the fruit fly. J. Neurosci. 2005;25:7377–7384. PubMed PMC

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680–685. PubMed

Lawal HO, Chang H-Y, Terrell AN, Brooks ES, Pulido D, Simon AF, Krantz DE. The Drosophila vesicular monoamine transporter reduces pesticide-induced loss of dopaminergic neurons. Neurobiol. Dis. 2010;40:102–112. PubMed PMC

Mackay W, Reynolds ER, O'Donnell JM. Tissue-specific and complex complementation patterns in the Punch locus of Drosophila melanogaster. Genetics. 1985;111:885–904. PubMed PMC

Makos MA, Kim YC, Han KA, Heien ML, Ewing AG. In vivo electrochemical measurements of exogenously applied dopamine in Drosophila melanogaster. Anal. Chem. 2009;81:1848–1854. PubMed PMC

Mustard JA, Pham PM, Smith BH. Modulation of motor behavior by dopamine and the D1-like dopamine receptor AmDOP2 in the honey bee. J. Insect Physiol. 2010;56:422–430. PubMed PMC

Neckameyer WS, White K. Drosophila tyrosine hydroxylase is encoded by the pale locus. J. Neurogenet. 1993;8:189–199. PubMed

Neckameyer WS, Woodrome S, Holt B, Mayer A. Dopamine and senescence in Drosophila melanogaster. Neurobiol. Aging. 2000;21:145–152. PubMed

Pendleton RG, Rasheed A, Sardina T, Tully T, Hillman R. Effects of tyrosine hydroxylase mutants on locomotor activity in Drosophila: a study in functional genomics. Behav. Genet. 2002;32:89–94. PubMed

Pfeiffenberger C, Lear BC, Keegan KP, Allada R. Locomotor activity level monitoring using the Drosophila Activity Monitoring (DAM) System. Cold Spring Harb. Protoc. 2010;11:pdb.prot5518. PubMed

Porzgen P, Park SK, Hirsh J, Sonders MS, Amara SG. The antidepressant-sensitive dopamine transporter in Drosophila melanogaster: a primordial carrier for catecholamines. Mol. Pharmacol. 2001;59:83–95. PubMed

Rauschenbach I.Yu., Karpova EK, Adonyeva NV, Andreenkova OV, Faddeeva NV, Burdina EV, Alekseev AA, Menshanov PN, Gruntenko NE. Disruption of insulin signaling affects the neuroendocrine stress reaction in Drosophila females. J.Exp. Biol. 2014 (In Press: doi: 10.1242/?jeb.106815) PubMed

Ream PJ, Suljak SW, Ewing AG, Han K-A. Micellar electro-kinetic capillary chromatography-electrochemical detection for analysis of biogenic amines in Drosophila melanogaster. Anal. Chem. 2003;75:3972–3978. PubMed

Restifo L, White K. Molecular and genetic approaches to neurotransmitter and neuromodulator systems in Drosophila. Adv. Insect Physiol. 1990;22:116–219.

Reynolds ER, O'Donnell JM. Characterization of new Punch mutations: identification of two additional mutant classes. Genetics. 1988;119:609–617. PubMed PMC

Romero-Calderon R, Uhlenbrock G, Borycz J, Simon AF, Grygoruk A, Yee SK, Shyer A, Ackerson LC, Maidment NT, Meinertzhagen IA, Hovemann. B.T. Krantz, D.E. A glial variant of the vesicular monoamine transporter is required to store histamine in the Drosophila visual system. PLoS Genet. 2008;4:e1000245. PubMed PMC

Rushmore TH, Morton MR, Pickett CB. The antioxidant responsive element. Activation by oxidative stress and identification of the DNA consensus sequence required for functional activity. J. Biol. Chem. 1991;266:11632–11639. PubMed

Rushmore TH, Pickett CB. Transcriptional regulation of the rat glutathione S-transferase Ya subunit gene. Characterization of a xenobiotic-responsive element controlling inducible expression by phenolic antioxidants. J. Biol. Chem. 1990;265:14648–14653. PubMed

Salinas AE, Wong MG. Glutathione S-Transferases - A Review. Curr. Med. Chem. 1999;6:279–309. PubMed

Simon AF, Daniels R, Romero-Calderon R, Grygoruk A, Chang H-Y, Najibi R, Shamouelian D, Salazar E, Solomon M, Ackerson LC, Maidment NT, DiAntonio A, Krantz DE. Drosophila vesicular monoamine transporter mutants can adapt to reduced or eliminated vesicular stores of dopamine and serotonin. Genetics. 2009;181:525–541. PubMed PMC

Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC. Measurement of protein using bicinchonininc acid. Anal. Biochem. 1985;150:76–85. PubMed

Stathakis DG, Burton Y, McIvor WE, Krishnakumar S, Wright TRF, O'Donnell JM. The catecholamines up (Catsup) protein of Drosophila melanogaster functions as a negative regulator of tyrosine hydroxylase activity. Genetics. 1999;153:361–382. PubMed PMC

Ueda S, Aikawa M, Ishizuya-Oka A, Yamaoka S, Koibuchi N, Yoshimoto K. Age-related dopamine deficiency in the mesostriatal dopamine system of zitter mutant rats: regional fiber vulnerability in the striatum and the olfactory tubercle. Neuroscience. 2000;95:389–398. PubMed

Ueno T, Masuda N, Kume S, Kume K. Dopamine modulates the rest period length without perturbation of its power law distribution in Drosophila melanogaster. PloS One. 2012;7:e32007. PubMed PMC

Waddell S. Reinforcement signaling in Drosophila; dopamine does it all after all. Curr. Opin. Neurobiol. 2013;23:324–329. PubMed PMC

Wang Z, Ferdousy F, Lawal H, Huang Z, Daigle JG, Izevbaye I, Doherty O, Thomas J, Stathakis DG, O'Donnell JM. Catecholamines up integrates dopamine synthesis and synaptic trafficking. J. Neurochem. 2011;119:1294–1305. PubMed PMC

Wulff K, Gatti S, Wettstein JG, Foster RG. Sleep and circadian rhythm disruption in psychiatric and neurodegenerative disease. Nat. Rev. Neurosci. 2010;11:589–599. PubMed

Yujnovsky I, Hirayama. Dol, M., Borrelli E, Sassone-Corsi P. Signaling mediated by the dopamine D2 receptor potentiates circadian regulation by CLOCK:BMAL1. Proc. Natl. Acad. Sci. USA. 2006;103:6386–6391. PubMed PMC

Elimination of certain honeybee venom activities by adipokinetic hormone

Disruption of dopamine homeostasis has sexually dimorphic effects on senescence characteristics of Drosophila melanogaster