Transcription activity of genes taking part in Colorado potato beetle diapause regulation and its changes under the fipronil impact

Cover Page

Abstract


Background. The peculiarities of transcription activity of genes taking part in Colorado potato beetle diapause regulation have been investigated under the laboratory conditions in winter diapause initiation phase and in the middle of winter. 

Materials and Methods. Excerpt of summer generation (2015 year.) adults divided in active and inactive individuals in groups treated or untreated by fipronil in sublethal concentration (0.0001% a.i.) in the diapause initiation and maintenance phases. Quantitative real-time PCR used for comparing evaluation of transcription activity in muscles and gonads tissues for genes DP1, fer, DAT1, DAT2, EcR, Ldace-1 and rdl. 
Results. In gonads of active adults unlike the quiescent ones detected presence of rdl subunit of GABA-А receptor mRNA. In the middle of diapause maintenance phase in fipronil-treated adults revealed the increasing of mRNA content in muscles and gonads for genes of reserve proteins DP1, defensive proteins DAT1, DAT2, ferritin fer, gene of ecdysone receptor EcR and gene coding the acetylcholinesterase Ldace-1. 

Conclusion. The assumption advanced about the fipronil blocking the GABA-receptor binding thereby has significant impact to regulation processes accompanying the development of diapause states.


Yuri M. Nikonorov

Ufa Scientific Center of RAS

Author for correspondence.
Email: nikonorov@anrb.ru

Russian Federation Senior researcher, Laboratory of molecular biology and nanobiotechnology. Institute of Biochemistry and Genetic

Liana A. Syrtlanova

Ufa Scientific Center of RAS

Email: slian4ik@mail.ru

Russian Federation Post-graduate student, Laboratory of physiological genetics. Institute of Biochemistry and Genetics

Konstantin A. Kitaev

FGBU Bashkirskiy referentniy centr Rosselhoznadzora

Email: cordek@ya.ru

Russian Federation Entomologist, Laboratory of phytosanitary research

Galina V. Benkovskaya

Ufa Scientific Center of RAS

Email: bengal2@yandex.ru

Russian Federation Leading researcher, Laboratory of physiological genetics. Institute of Biochemistry and Genetics

  1. Lehman P, Piironen S, Kankaret M, et al. Photoperiodic effects on diapause-associated gene expression trajectories in European Leptinotarsa decemlineata populations. Insect Mol Biol. 2014;23(5):566-578. doi: 10.1111/jmb.12104.
  2. Саулич А.Х. Сезонное развитие насекомых и возможности их расселения. — СПб.: Изд-во СПбГУ, 1999. — 248 с. [Saulich AH. The seasonal development of insects and the possibility of resettlement. Saint Petersburg: Izdatelstvo SpbGU; 1999: 248 p. (In Russ.)]
  3. Yocum GD, Rinehart JP, Larson ML. Monitoring diapause development in the Colorado potato beetle, Leptinotarsa decemlineata, under field conditions using molecular biomarkers. J Insect Physiol. 2011;57(5):645-652. doi: 10.1016/j.jinsphys.2010.11.008.
  4. Denlinger DL. Regulation of diapause. Ann Rev Entomol. 2002;47:93-122. doi: 10.1146/annurev.ento.
  5. 091201.145137.
  6. Беньковская Г.В. Стресс-реакция как механизм реализации адаптивного потенциала особей и популяций насекомых: Дис. … д-ра биол. наук. — Уфа: ИБГ УНЦ РАН, 2008. [Benkovskaya GV. Stress reaction as a mechanism of realization for adaptive potential of individuals and populations of insects. [dissertation]. Ufa: IBG UCS RAS; 2008. (In Russ.)]
  7. Yocum GD, Rinehart JP, Chirumamilla-Chapara A, Larson ML. Characterization of gene expression patterns during the initiation and maintenance phases of diapause in the Colorado potato beetle, Leptinotarsa decemlineata. J Insect Physiol. 2009;55:32-39. doi: 10.1016/j.jinsphys.2008.10.003.
  8. Enell L, Hamasaka Y, Kolodziejczyk A, Nässel DR. Gamma-Aminobutyric acid (GABA) signaling components in Drosophila: immunocytochemical localization of GABA(B) receptors in relation to the GABA(A) receptor subunit RDL and a vesicular GABA transporter. J Comp Neurol. 2007;505(1):18-31. doi: 10.1002/cne.21472.
  9. Pham DQD, Winzerling JJ. Insect ferritins: typical or atypical? Biochim Biophys Acta. 2010;1800(8):824-833. doi: 10.1016/j.bbagen.2010.03.004.
  10. Shi XQ, Guo WC, Wan PJ, et al. Validation of reference genes for expression analysis by quantitative real-time PCR in Leptinotarsa decemlineata (Say). BMC Research Notes. 2013;6:93. doi: 10.1186/1756–0500–6–93.
  11. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-∆∆CT method. METHODS. 2001;25:402-408. doi: 10.1006/meth.2001.1262.
  12. Беньковская Г.В., Мустафина Р.Ш. Влияние светового режима на биохимические показатели развития стресс-реакции в линиях Musca domestica L. с различной продолжительностью жизни // Журнал эволюционной биохимии и физиологии. — 2012. — Т. 48. — № 5. — С. 433–438. [Benkovskaya GV, Mustafina RSh. Effect of light regime on biochemical parameters of development of stress-reactions of Musca domestica lines with different lifespan. Zhurnal evolyutsionnoi biohimii i physiologii. 2012;48(5):433-438. (In Russ., Eng.)]. doi: 10.1134/S0022093012050027.
  13. Planello R, Martinez-Guitarte JL, Morcillo G. The endocrine disruptor bisphenol A increases the expression of Hsp70 and ecdysone receptor genes in the aquatic larvae of Chironomus riparius.Chemosphere. 2008;71(10):1870-1876.
  14. doi: 10.1016/j.chemosphere.2008.01.033.
  15. Planello R, Herrero O, Martinez-Guitarte JL, Morcillo G. Comparative effect of butyl benzyl phthalate (BBP) and di(2-ethylhexyl) phthalate (DEHP) on the aquatic larvae of Chironomus riparius based on gene expression assays related to the endocrine system, the stress response and ribosomes. Aquat Toxicol. 2011;105(1-2):62-70.
  16. doi: 10.1016/j.aquatox.2011.05.011.
  17. Buckingham SD, Biggin PC, Sattelle BM, et al. Insect GABA receptors: splicing, editing, and targeting by antiparasitics and insecticides. Mol Pharmacol. 2005;68:942-951. doi: 10.1124/mol.105.015313.
  18. Wang RL, Bencic D, Villeneuve DL, et al. A transcriptome-based biological framework for studying mechanisms of endocrine disruption in small fish species. Aquat Toxicol. 2010;98(3):230-244. doi: 10.1016/j.aquatox.2010.02.021.
  19. Roques BB, Leghait J, Lacroix MZ, et al. The nuclear receptors pregnane X receptor and constitutive androstane receptor contribute to the impact of fipronil on hepatic gene expression linked to thyroid hormone metabolism. Biochem Pharmacol. 2013;86(7):997-1039. doi: 10.1016/j.bcp.2013.08.012.
  20. Тыщенко В.П., Кинд Т.В. Нейроэндокринные механизмы регуляции сезонных циклов // Гормональная регуляция развития насекомых. — Л.: Наука, 1983. — Глава 5. — С. 82–117. [Tyshchenko VP, Kind TV. Neuroendocrine mechanisms of regulation of seasonal cycles. Leningrad: Nauka. Chapter 5, Hormonal regulation of insect development; 1983. P. 82-117.
  21. Kubrak OI, Kučerova I, Theopold U, Nässel DR. The sleeping beauty: how reproductive diapause affects hormone signaling, metabolism, immune response and somatic maintenance in drosophila melanogaster. PLoS One. 2014;9(11):e 113051. doi: 10.1371/journal. pone. 0113051.
  22. Kumar A, Congiu L, Lindström L, et al. Sequencing, De Novo assembly and annotation of the Colorado potato beetle, Leptinotarsa decemlineata, transcriptome. PLoS One. 2014;9(1): e86012. doi: 10.1371/journal. pone. 0086012.
  23. Xu WH, Lu YX, Denlinger DL. Cross-talk between the fat body and brain regulates insect developmental arrest. PNAS. 2012;109(36). doi: 10.1073/pnas.1212879109.
  24. Ehlen JC, Paul KN. Regulation of light’s action in the mammalian circadian clock: role of the extrasynaptic GABAA receptor. Am J Physiol Regul Integr Comp Physiol. 2009;296(5):1606-1612. doi: 10.1152/ajpregu.90878.2008.
  25. De Loof A. The essence of female-male physiological dimorphism: differential Ca2+-homeostasis enabled by the interplay between farnesol-like endogenous sesquiterpenoids and sex-steroids? The calcigender paradigm. Gen Comp Endocrinol. 2015;211:131-146. doi: 10.1016/j.ygcen.2014.12.003.
  26. Wei Q, Wu SF, Niu CD, et al. Knockdown of the ionotropic γ-aminobutyric acid receptor (GABAR) RDL gene decreases fipronil susceptibility of the small brown planthopper, Laodelphax striatellus(Hemiptera: Delphacidae). Arch Insect Biochem Physiol. 2015;88(4): 249-261. doi: 10.1002/arch.21232.
  27. Taylor-Wells J, Brooke BD, Bermudez I, Jones AK. The neonicotinoid imidacloprid and the pyrethroid deltamethrin are antagonists of the insect Rdl GABA receptor. J Neurochem. 2015;135(4):705-7013. doi: 10.1111/jnc.13290.
  28. Impagnatiello F, Pesold C, Longone P, et al. Modifications of gamma-aminobutyric acid A receptor subunit expression in rat neocortex during tolerance to diazepam. Mol Pharmacol. 1996;49(5):822-831. PMID: 8622632.
  29. Беньковская Г.В., Китаев К.А., Удалов М.Б. Успех переживания экспериментальной диапаузы и резистентность к инсектицидам в популяциях колорадского жука / Экология, эволюция и систематика животных: материалы международной научно-практической конференции. Ноябрь 13–16, 2012; Рязань, с. 45–46. [Benkovskaya GV, Udalov MB, Kitaev KA. The success of survival under experimental diapause and insecticide resistance in Colorado beetle populations in the South Urals. Ecologia, evoliyutsia I sistematika zhivotnyh [conference proceedings]. 2012 November 13-16. Ryazan. P. 45-46. (In Russ.)]

Supplementary files

Supplementary Files Action
1. Fig. 1. Differential expression in gonad and muscle tissues of Colorado potato beetle genes coding the diapause-associated proteins (DP1, DAT1, DAT2, DAT3) and gene of ecdysone receptor (Ld EcR) in the phase of diapause initiation (* — significant difference between active and inactive individuals, Mann-Whitney test, p = 0.01). On Y axis — ratio of mRNA copy number in gonads toward mRNA copy number in muscles, referent gene — rp4 View (86KB) Indexing metadata
2. Fig. 2. Сhanges of transcriptional activity under the fipronil impact: (а) diapause-associated proteins genes (DP1, DAT1, DAT2, fer; * — significant difference between active and inactive individuals, Mann-Whitney test, p = 0.01); (b) gene of ecdysone receptor (Ld EcR) (* —significant difference between gonads and muscles, Mann-Whitney test, p = 0.01) in Colorado potato beetle adults in the phase of diapause maintenance View (101KB) Indexing metadata
3. Fig. 3. The activity of acetylcholine esterase in Colorado potato beetle adult’s haemolymph in winter reproductive diapause and its changes under the fipronil impact. * — significant difference between fipronil-treated and control individuals, Student’s test, р ≤ 0.01 View (32KB) Indexing metadata
4. Fig. 4. rdl subunit gene transcriptional activity in the gonad and muscle tissues of adult Colorado potato beetles. a: in the diapause initiation phase (* — significant difference between active and inactive individuals, Mann-Whitney test, p = 0.01); b: in the diapause maintenance, (* — significant difference between fipronil-treated and control individuals, Mann-Whitney test, p = 0.01), mRNA content ratio shown in fipronil treated and control individuals View (56KB) Indexing metadata

Views

Abstract - 1193

PDF (Russian) - 169

PlumX


Copyright (c) 2016 Nikonorov Y.M., Syrtlanova L.A., Kitaev K.A., Benkovskaya G.V.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.