Aleinikava N.Y.1, Boika A.V.1, Nizheharodava D.B.1, Ponomarev V.V.1, Vanslau M.I.1, Ustsiamchuk A.M.1, Ihnatovich T.V.1, Kuznetsova T.Y.2, Hladkova Z.A.2, Zafranskaya M.M.1
The obtaining of toxic chronic model of parkinsonism syndrome in rats
1Belarusian Medical Academy of Post-Graduate Education, Minsk, Republic of Belarus
2Institute of Physiology of the Belarusian National Academy of Sciences, Minsk, Republic of Belarus
Vestnik VGMU. 2018;17(6):92-99.
Introduction. The investigation of parkinsonian syndrome (PS) models in vivo and in vitro allows to indirectly judge about the processes taking place in the living human cells. Neurotoxins are used for obtaining selective death of black substance (BS) neurons in PS modelling.
Objectives. To create chronic toxic experimental morphologically confirmed model of PS in rats on systemic administration of rotenone.
Material and methods. White mongrel female rats weighing 250-350 g (n=80) were used. The following solvent was made use of to dissolve rotenone – 99% dimethylsulfoxide (DMSO):20% Lipovenos, 1:1. Rotenone was administered subcutaneously in the rostral parts of the interscapular area at the doses of 1,75 mg / kg and 2,0 mg / kg (main group, n=60). The control group (n=20) consisted of animals, which under the same conditions were injected with the solvent in an equivalent volume. The individual identification of rats was used. Frontal sections of the brain with a thickness of 7μm were prepared on the microtome-cryostat HM 525. For the light-optical examination, the sections were stained with thionine and methylene blue according to Nissl and hematoxylin-eosin. Altami LUM-1 microscope with a digital camera and software with an 40x lens increase was used. Results. In the main group, the clinical symptoms characteristic of experimental PS developed in 3 days after the introduction of rotenone. In the control group, there were no clinical signs of PS. After the introduction of the solvent, no significant structural changes in the neurons of the rats’ black substance were detected. In the main group in the black brain substance of rats, after the initial daily administration of rotenone at the dose of 1,75 and 2 mg / kg, pronounced destructive changes were observed. Microscopy revealed in the black substance a considerable number of cells with destructive and dystrophic changes, in some neurons of the black substance Lewy bodies were also found. Conclusions. It is necessary to satisfy all the requirements on the quality of the experiment being conducted to obtain chronic neurotoxic model of PS, reflecting the classical symptomatology of PD.
Key words: Parkinson’s disease, modelling on rats, rotenone, morphology, Lewy bodies.
1. Voronina EA, Val'dman LN, Nerobkova, Kapitsa IG. Guidelines for preclinical study of drugs with anti-Parkinsonian activity. V: Seredenin SB, Khaitov RM, Anokhina IP, Archakov AI, Ramnbykov VA, Gintsburg AL, i dr. Rukovodstvo po provedeniiu doklinicheskikh issledovanii lekarstvennykh sredstv Ch 1. Moscow, RF: Grif i K; 2012. Gl 13. Р. 219-34. (In Russ.)
2. Caboni P, Sherer TB, Zhang N, Taylor G, Na HM, Greenamyre JT, et al. Rotenone, deguelin, their metabolites, and the rat model of Parkinson's disease. Chem Res Toxicol. 2004 Nov;17(11):1540-8.
3. Malinovskaya NA, Morozova GA, Kuvacheva NV, Gasymly ED. Models of Parkinson's disease in vitro. Nauch Aspekt. 2012;2(4):193-201. (In Russ.)
4. Betarbet R, Sherer TB, Greenamyre JT. Animal models of Parkinson Disease. Bioessays. 2002 Apr;24(4):308-18. doi: http://dx.doi.org/10.1002/bies.10067
5. Illarioshkin SN, Khaspekov LG, Grivennikov IA. Simulation of Parkinson's disease using induced pluripotent stem cells. Moscow, RF: RKI Sovero press; 2016. 183 р. (In Russ.)
6. Ricaurte GA, Langston JW, Delanney LE, Irwin I, Peroutka SJ, Forno LS. Fate of nigrostriatal neurons in young mature mice given 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine: a neurochemical and morphological reassessment. Brain Res. 1986 Jun;376(1):117-24.
7. Jackson-Lewis V, Jakowec M, Burke RE, Przedborski S. Time course and morphology of dopaminergic neuronal death caused by the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Neurodegeneration. 1995 Sep;4(3):257-69.
8. Panov A, Dikalov S, Shalbuyeva N, Taylor G, Sherer T, Greenamyre JT. Rotenone Model of Parkinson Disease. Multiple brain mitochondria dysfunctions after short term systemic rotenone intoxication. J Biol Chem. 2005 Dec;280(51):42026-35. doi: http://dx.doi.org/10.1074/jbc.M508628200
9. Anisimov SV. Cell therapy for Parkinson's diseasestem cells. Saint-Petersburg, RF: Izd-vo N-L; 2014. 320 р. (In Russ.)
10. Thiffault C, Langston JW, Di Monte DA. Increased striatal dopamine turnover following acute administration of rotenone to mice. Brain Res. 2000 Dec;885(5):283-8.
11. Betarbet R, Sherer TB, MacKenzie G, Garcia-Osuna M, Panov AV, Greenamyre JT. Chronic systemic pesticide exposure reproduces features of Parkinson's disease. Nat Neurosci. 2000 Dec;3(12):1301-6. doi: http://dx.doi.org/10.1038/81834
12. Sherer TB, Kim JH, Betarbet R, Greenamyre JT. Subcutaneous rotenone exposure causes highly selective dopaminergic degeneration and alpha-synuclein aggregation. Exp Neurol. 2003 Jan;179(1):9-16.
Information about authors:
Aleinikava N.Y. – postgraduate of the Chair of Neurology & Neurosurgery, Belarusian Medical Academy of Post-Graduate Education;
Boika A.V. – Candidate of Medical Sciences, doctoral candidate of the Chair of Neurology & Neurosurgery, Belarusian Medical Academy of Post-Graduate Education;
Nizheharodava D.B. – Candidate of Biological Sciences, leading research officer of the immunology and biomedical technologies department of the Scientific-Research Laboratory, Belarusian Medical Academy of Post-Graduate Education;
Ponomarev V.V. – Doctor of Medical Sciences, professor, head of the Chair of Neurology & Neurosurgery, Belarusian Medical Academy of Post-Graduate Education;
Vanslau M.I. – associate research officer of the immunology and biomedical technologies department of the Scientific-Research Laboratory, Belarusian Medical Academy of Post-Graduate Education;
Ustsiamchuk A.M. – associate research officer of the immunology and biomedical technologies department of the Scientific-Research Laboratory, Belarusian Medical Academy of Post-Graduate Education;
Ihnatovich T.V. – associate research officer of the Scientific-Research Laboratory, Belarusian Medical Academy of Post-Graduate Education;
Kuznetsova T.Y. – Candidate of Biological Sciences, senior research officer of the Institute of Physiology, Belarusian National Academy of Sciences;
Hladkova Z.A. – research officer of the Neurophysiology Laboratory of the Institute of Physiology, Belarusian National Academy of Sciences;
Zafranskaya M.M. – Doctor of Medical Sciences, principal research officer of the immunology and biomedical technologies department of the Scientific-Research Laboratory, Belarusian Medical Academy of Post-Graduate Education.