Effect of novelty stress on behavioral responses of Danio rerio and assessment of dose-dependent effects of anxiolytics of benzodiazepine structure with phenazepam as an example

Cover Page

Abstract


The effect of benzodiazepine anxiolytic phenazepam in Danio rerio was investigated. Previously, Danio rerio showed the effects of other anxiolytics, dia_zepam and chlordiazepoxide. The analysis of the anxiolytic effect of phenazepam in Danio rerio was carried out for the first time.

Methods. A stress test on novelty situation was used: a fish was placed first in a beaker with a dissolved pharmacological substance (or water) and then into a novel tank for 6 min, where the trajectory, the path length, the number of movements to the upper part of the novel tank, the number and time of the pattern of “freezing” for each min of the experiment were measured.

Results. In response to the novelty of tank, the fish was shown to react by submerging to the bottom, increasing the freezing, and reducing the number of movements to the upper half of the novel tank. After phenazepam exposure (administration), the fish was not only in the lower, but also in the upper part of the novel tank. The average path length did not change significantly in the range of the doses used. The number and time of the freezing, as well as the time spent in the lower part of the novel tank, 2-fold decreased compared to the control group of animals and showed a dose-dependent effect. The number of movements to the upper part of the novel tank for the experiment increased significantly from 1 in the control to 57 after phenazepam in a dose of 1 mg/l. When analyzing the dynamics of the parameters for each min, it was shown that the time of the fish in the lower part of the novel tank decreased from 3th min of the experiment with the use of phenazepam in a dose of 0.5 mg/l. At the same time, the number of movements of fish to the upper part of the novel tank significantly increased more than 2 times from 3th min of the experiment with the use of phenazepam in a dose of 1 mg/l.

Conclusion. The described method to study behavioral responses of Danio rerio on novelty stress is high sensitive in comparison with traditional behavioral methods of studing tranquilizers. The prospect of using Danio rerio as animal model in behavioral pharmacology is significant and does not concede research on rodents.


Petr D. Shabanov

Author for correspondence.
pdshabanov@mail.ru
Institute of Experimental Medicine
Russian Federation, 12, Academic Pavlov street, Saint-Petersburg, 197376

Dr Med Sci, Professor, Head, S.V. Anichkov Dept. of Neuropharmacology

Viktor A. Lebedev

aalebedev-iem@rambler.ru
Institute of Experimental Medicine
Russian Federation, 12, Academic Pavlov street, Saint-Petersburg, 197376

PhD (Pharmacology), Ingeneer, S.V. Anichkov Dept. of Neuropharmacology

Andrei A. Lebedev

aalebedev-iem@rambler.ru
Institute of Experimental Medicine
Russian Federation, 12, Academic Pavlov street, Saint-Petersburg, 197376

Dr Biol Sci (Pharmacology), Leading Researcher, S.V. Anichkov Dept. of Neuropharmacology

Eugenii R. Bychkov

bychkov@mail.ru
Institute of Experimental Medicine
Russian Federation, 12, Academic Pavlov street, Saint-Petersburg, 197376

PhD (Biochemistry), Leading Researcher, S.V. Anichkov Dept. of Neuropharmacology

  • Лебедев А.А., Пшеничная А.Г., Бычков Е.Р., и др. Антагонист рецепторов кортиколиберина астрессин снимает тревожно-фобические состояния у крыс, выращенных в социальной изоляции // Обзоры по клинической фармакологии и лекарственной терапии. – 2016. – Т. 14. – № 4. – С. 24–31. [Lebedev AA, Pshenichnaya AG, Bychkov ER, et al. Antagonist receptorov kortikoliberina astressin snimaet trevozhno-fobicheskie sostoyaniya u krys, vyrashchennyh v social’noj izolyacii. Review on Clinical Pharmacology and Medicinal Therapy. 2016;14(4):24-31. (In Russ.)]. doi: 10.17816/RCF14424-31.
  • Шабанов П.Д., Лебедев А.А., Якушина Н.Д., и др. Моделирование обсессивно-компульсивного и аддиктивного игрового поведения у крыс введением фенамина в тесте закапывания шариков // Наркология. – 2017. – Т. 16. – № 1. – С. 32–38. [Shabanov PD, Lebedev AA, Yakushina ND, et al. Modelirovanie obsessivno-kompul’sivnogo i addiktivnogo igrovogo povedeniya u krys vvedeniem fenamina v teste zakapyvaniya sharikov. Narkologiya. 2017;16(1):32-38. (In Russ.)]
  • Bencan Z, Sledge D, Levin ED. Buspirone, chlordiazepoxide and diazepam effects in a Danio rerio model of anxiety. Pharmacol Biochem Behav. 2009;94:75-80. doi: 10.1016/j.pbb.2009.07.009.
  • Best JD, Alderton WK. Danio rerio: an in vivo model for the study of neurological diseases. Neuropsychiatr Dis Treat. 2008;4:567-576.
  • Best JD. Non-associative learning in larval Danio rerio. Neuropsychopharmacology. 2008;33:1206-1215. doi: 10.1038/sj.npp.1301489.
  • Blaser R, Gerlai R. Behavioral phenotyping in Danio rerio: comparison of three behavioral quantification methods. Behav Res Methods. 2006;38:456-469. doi: 10.3758/BF03192800.
  • Bretaud S. A choice behavior for morphine reveals experience-dependent drug preference and underlying neural substrates in developing larval Danio rerio. Neuroscience. 2007;146:1109-1116. doi: 10.1016/j.neuroscience.2006.12.073.
  • Bronikowski AM. Open-field behavior of house mice selectively bred for high voluntary wheel-running. Behav Genet. 2001;31:309-316. doi: 10.1023/A:1012283426530.
  • Cachat J, Stewart A, Grossman L, Kalueff AV. Measuring behavioral and endocrine responses to novelty stress in adult Danio rerio. Nature protocols. 2010;5(11):1786-9. doi: 10.1038/nprot.
  • Choleris E, Thomas AW, Kavaliers M, Prato FS. A detailed ethological analysis of the mouse open field test: effects of diazepam, chlordiazepoxide and an extremely low frequency pulsed magnetic field. Neurosci Biobehav Rev. 2001;25:235-260. doi: 10.1016/S0149-7634(01)00011-2.
  • Creton R. Automated analysis of behavior in Danio rerio larvae. Behav Brain Res. 2009;203:127-136. doi: 10.1016/j.bbr.2009.04.030.
  • Egan RJ. Understanding behavioral and physiological phenotypes of stress and anxiety in Danio rerio. Behav Brain Res. 2009;205:38-44. doi: 10.1016/j.bbr.
  • Gerlai R. Event recording and video-tracking: towards the development of high-throughput Danio rerio screens. In: Proc. 5th Conf. on Methods in Behav. Res. Eds. J.J. Noldus, F. Grieco, L.W.S. Loijens, P.H. Zimmerman. 2005.
  • Gerlai R, Lee V, Blaser R. Effects of acute and chronic ethanol exposure on the behavior of adult Danio rerio (Danio rerio). Pharmacol Biochem Behav. 2006;85:752-761. doi: 10.1016/j.pbb.2006.11.010.
  • Guo S. Using Danio rerio to assess the impact of drugs on neural development and function. Expert Opin Drug Discov. 2009;4:715-726. doi: 10.1517/17460440902988464.
  • Key B, Devin CA. Danio rerio as an experimental model: strategies for developmental and molecular neurobiology studies. Methods Cell Sci. 2003;25:1-6. doi: 10.1023/B:. MICS.0000006849.98007.03.
  • Levin ED, Chen E. Nicotinic involvement in memory function in Danio rerio. Neurotoxicol Teratol. 2004;26:731-5. doi: 10.1016/j.ntt.2004.06.010.
  • Levin ED, Bencan Z, Cerutti DT. Anxiolytic effects of nicotine in Danio rerio. Physiol Behav. 2007;90:54-58. doi: 10.1016/j.physbeh.2006.08.026.
  • Lopez-Patino MA, Cabral H, Zhdanova IV. Anxiogenic effects of cocaine withdrawal in Danio rerio. Physiol Behav. 2008;93:160-171. doi: 10.1016/j.physbeh.2007.08.013.
  • Maximino C, Marques BT, Dias CA, et al. Parametric analyses of anxiety in Danio rerio scototaxis. Behav Brain Res. 2010;210:1-7. doi: 10.1016/j.bbr.2010.01.031.
  • Maximino C, Marques BT, Dias CA, et al. Scototaxis as anxiety-like behavior in fish. Nat Protoc. 2010;5:209-216. doi: 10.1038/nprot.2009.225.
  • Miklosi A, Andrew RJ. The Danio rerio as a model for behavioral studies. Danio rerio. 2006;3:227-234. doi: 10.1089/zeb.2006.3.227.
  • Ninkovic J, Bally-Cuif L. The Danio rerio as a model system for assessing the reinforcing properties of drugs of abuse. Methods. 2006;39:262-274. doi: 10.1016/j.ymeth.2005.12.007.
  • Sackerman J. Danio rerio behavior in novel environments: effects of acute exposure to anxiolytic compounds and choice of Danio rerio line. Int J Comp Psychol. 2010;23:43-61.
  • Spence R, Gerlach G, Lawrence C, Smith C. The behaviour and ecology of the Danio rerio. Biol Rev Camb Philos Soc. 2008;83:13-34. doi: 10.1111/j.1469-185X.2007.00030.x.
  • Stewart A. The developing utility of Danio rerio in modeling neurobehavioral disorders. Int J Comp Psychol. 2010;23:104-121. doi: 10.1016/j.pnpbp.2010.11.035.
  • Streng J. Open-field behavior in four inbred mouse strains. Can J Psychol. 1971;25:62-68. doi: 10.1037/h0082368.
  • Wong K. Analyzing habituation responses to novelty in Danio rerio (Danio rerio). Behav Brain Res. 2009;208:457. doi: 10.1016/j.bbr.2009.12.023.

Views

Abstract - 109

PDF (Russian) - 94


Copyright (c) 2017 ECO-vector LLC

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