Molecular Cloning and Expression Analysis of TH Gene in the Ant Polyrhachis vicina (Hymenoptera: Formicidae)

Gengsi Xi


Tyrosine hydroxylase (TH) plays an important role in the synthesis of L-DOPA. In this report, a TH homologue was isolated from Polyrhachis vicina Roger (Hymenoptera: Formicidae). The full –length cDNA of PvTH is 2429bp, which contains an open reading frame of 1713 bp. The 5’-and 3’- UTR s are 357 and 359 bp long, respectively. The open reading frame encodes a deduced 570 amino-acid peptide with a calculated molecular mass of 65.09 kilodaltons and an isoelectric point of 5.26. To compare PvTH mRNA expression during P. vicina development and the heads of different castes, real-time quantitative reverse transcription polymerase chain reaction was used. The results show that PvTH mRNA is differentially expressed in the ant development, in whole bodies and in the heads of different castes. During the development, the highest expression level is in pupae. The levels also vary among castes, the highest level is in workers. The investigation revel that PvTH is related with the developmental and caste-specifically at the level of transcription. 


tyrosine hydroxylase (TH); molecular cloning; Polyrhachis vicina Roger; real-time quantitative PCR

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Andretic, R., B. van Swinderen & R.J. Greenspan 2005. Dopaminergic modulation of arousal in Drosophila. Current Biology, 15:1165–1175.

Bendtsen, J.D., H. Nielsen, G. Von Heijine & S. Brunak 2004. Improved prediction of signal peptides: SignalP 3.0. Journal of Biochemistry and Molecular Biology, 340:783-795.

Birman, S., B. Morgan, M. Anzivino & J. Hirsh 1994. A novel and major isoform of tyrosine hydroxylase in Drosophila is generated by alternative RNA processing. Journal of Biological Chemistry, 269:26559–26567.

Budnik, V. & K.White 1988. Catecholamine-containing neurons in Drosophila melanogaster: distribution and development. Journal of Comparative Neurology, 268:400–413.

Chang H.Y., A. Grygoruk, E.S. Brooks, L.C. Ackerson, N.T. Maidment, R.J. Bainton & D.E. Krantz 2006. Overexpression of the Drosophila vesicular monoamine transporter increases motor activity and courtship but decreases the behavioral response to cocaine. Molecular Psychiatry, 11:99–113.

Fitzgerald, M. & T. Shenk 1981. The sequence 5’-AAUAAA-3’ forms parts of the recognition site for polyadenylation of late: SV40 mRNAs. Cell, 24: 251-260.

Futahashi, R. & H. Fujiwara 2005. Melanin-synthesis enzymes coregulates stage-specific larval cuticular markings in the swallowtail butterfly, Papilio xuthus. Development Genes and Evolution, 215:519–529.

Futahashi, R., J. Sato & Y. Meng 2009. Yellow and ebony are the responsible genes for the larval color mutants of the silkworm. Genetics, 180: 1995–2005.

Ganguly-Fitzgerald I., J. Donlea & P.J. Shaw 2006. Waking experience affects sleep need in Drosophila. Science, 313:1775–1781.

Goldstein, M. & A. Deutch 1992. Dopaminergicmechanisms in the pathogenesis of schizophrenia. FASEB Journal, 6:2413–2421.

Gotz, P. & H.G. Boman 1985. Insect immunity. Comprehensive Insect Physiology,Biochemistry and Pharmacology, vol. 11. Pergamon Press, New York, pp. 453–485.

Haavik, J. & T. Flatmark 1987. Isolation and characterization of tetrahydropterin oxidation products generated in the tyrosine 3-monooxygenase (tyrosine hydroxylase) reaction, European Journal of Biochemistry, 168: 21–26.

Hopkins, T.L. & K.J. Kramer 1992. Insect cuticle sclerotization. Annual Review of Entomology, 37:273–302.

Ichikawa, S. H. Ichinose & T. Nagatsu 1990. Multiple mRNA as of monkey tyrosine hydroxylase. Biochemical and Biophysical Research Communications, 73:1311-1336.

Jeanmougin, F., J.D. Thompson, M. Gouy, D.G. Higgins, T.J. & Gibson 1998. Multiple sequence alignment with Clustal X. Trends in Biochemical Sciences, 23:403-405.

Kaneda, N., K. Kobayashi & H.Ichinose 1987. Isolation of Novel cDNA clone for humantyrosine hydroxylase: alternative RNA splicing produces four kinds of mRNA from a single gene. Biochemical and Biophysical Research Communications, 146:971-975.

Kobayashi, K. & S. Morita 1995. Targeted disruption of the tyrosine hydroxylase locus results in severe catecholamine depletion and perinatal lethality in mice. Journal of Biological Chemistry, 270:27235–27243.

Kume K., S. Kume S.K. Park, J.Hirsh & F.R. Jackson 2005. Dopamine is a regulator of arousal in the fruit fly. Journal of Neuroscience, 25:7377–7384.

Lu, S.M., G.S. Xi & X.H. Wang 2008. Molecular cloning, characterization, and expression analysis of a QM homologue in the ant Polyrhachis vicina (Hymenoptera:Formicidae). The Canadian Entomologist, 140(3):312-323.

Lundell M.J. & J.Hirsh 1994. Temporal and spatial development of serotonin and dopamine neurons in the Drosophila CNS. Developmental Biology165:385–396.

Matthias, V. & W. Christian 2008. Neuroarchitecture of Aminergic Systems in the Larval Ventral Ganglion of Drosophila melanogaster. Plos One, 3(3): e1848.

Meyer, M.M. & P.F. Fitzpatrick 1992. The amion acid substrate of bovine tyrosine hydroxylase. Neurochemistry International, 21:191-196.

Monastirioti, M. 1999. Biogenic amine systems in the fruit fly Drosophila melanogaster. Microscopy Research and Technique, 45:106–121.

Nagatsu, T., M. Levitt & S.Udenfriend 1964. Tyrosine hydroxylase. Journal of Biological Chemistry, 239:2910-2917.

Nässel D.R. & K.Elekes 1992. Aminergic neurons in the brain of blowflies and Drosophila: dopamine- and tyrosine hydroxylase-immunoreactive neurons and their relationship with putative histaminergic neurons. Cell and Tissue Research, 267:147–167.

Neckameyer W.S. & J.S.Weinstein 2005. Stress affects dopaminergic signaling pathways in Drosophila melanogaster. Stress, 8:117–131.

Ninomiya, Y. & Y. Hayakawa 2007. Insect cytokine, growth-blocking peptide, is a primary regulator of melanin-synthesis enzymes in armyworm larval cuticle. FEBS Journal, 274:1768-1777.

Ouyang, X.H., G.S. Xi & C.P. Bu 2009. Molecular cloning and expression of an estrogen receptor-related receptor gene in the ant Polyrhachis vicina (Hymenoptera:Formicidae). Annals of the Entomological Society of America, 102(2):295-302.

Peng, X.M. 2002. Tyrosine hydroxylase and Parkinson’s disease. Chinese Journal of Clinical Neurosciences, 10:105-108.

Restifo, L.L. & K. White 1990. Molecular and genetic approaches to neurotransmitter and neuromodulator systems in Drosophila. Advances in Insect Physiology, 22:116–219.

Schwaerzel M., M.Monastirioti, H.Scholz, F. Friggi-Grelin, S. Birman & M. Heisenberg 2003. Dopamine and octopamine differentiate between aversive and appetitive olfactory memories in Drosophila. Journal of Neuroscience, 23:10495–10502.

Söderhäll, K. & L.Cerenius 1998. Role of the prophenoloxidase-activating system in invertebrate immunity. Current Opinion in Immunology, 10:23–28.

SPSS Inc. 2004. SPSS . Version 13.0. SPSS Inc., Chicago.

Stathakis, D.G., D.Y. Burton, W.E. McIvor, S. Krishnakumar & T.R.F. Wright 1999. The catecholamines up (Catsup) protein of Drosophila melanogaster functions as a negative regulator of tyrosine hydroxylase activity. Genetics, 153:361–382.

Tamura, K., J. Dudley, M. Nei & S. Kumar 2007. MEGA4: molecular evolutionary genetics analysis (MEGA) software. Version 4.0. Molecular Biology and Evolution, 24:1596- 1599.

Truman, J.W. 1996. Steroid receptors and nervous system metamorphosis in insets. Developmental Neuroscience, 18:87-101.

Ulrich, R. & M. Hofrichter 2007. Enzymatic hydroxylation of aromatic compounds. Cellular and Molecular Life Sciences, pp. 271–293.

Wang Da-yong 2008. Molecular cloning, function analysis and prokaryotic expression of the tyrosine hydroxylase gene in mealworm, Tenebrio molitor (Insecta Coleoptera)

Ye Y., W. Xi, Y.Peng, Y.Wang & A. Guo 2004. Long-term but not short-term blockade of dopamine release in Drosophila impairs orientation during flight in a visual attention paradigm. European Journal of Neuroscience, 20:1001–1007.

Zhang, C.Y., S.G. Xu &X.X. Huang 2005. A novel and convenient relative quantitative method of fluorescence real time RT -PCR assay based on slope of standard curve. Progress in Biochemistry and Biophysiology, 32:883-888.



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