Localization of total and Ser 129 phosphorylated α-synuclein in the rat peripheral nervous system (testing of different antibodies)
Abstract
The deposition of α-synuclein (α-Syn) in the peripheral nervous system precedes the motor symptoms of Parkinson's disease and is considered as a pathological sign and biomarker. However, data on the localization of α-Syn in the intact nervous system require clarification for the future development of experimental models and diagnostic tests. Localization of α-Syn and its the remainder of serine 129 phosphorylated form in peripheral nervous system of intact rats were studied by immunohistochemistry with use of different commercially available antibodies. α-Syn were found in presynaptic terminals of spinal dorsal horn, neuronal somata of prevertebral ganglia, nerve plexuses of vascular wall and GFAP-negative glial cells. Different antibodies to phosphorylated α-Syn shown uneven binding. Staining of processes, somata or nuclei of certain dorsal root ganglia neurons were found. No one of used antibodies detected small nociceptive IB4-positive neurons. The ambiguity of the obtained results can be related both to the different availability of epitopes detected by antibodies as a result of α-Syn interactions with other proteins, and with non-specific binding of antibodies to unidentified targets.
About the Authors
D. N. Voronkov
ФГБНУ «Научный центр неврологии»
Russian Federation
Russian Federation
O. V. Salnikova
ФГБНУ «Научный центр неврологии»
Russian Federation
Russian Federation
R. M. Khudoerkov
ФГБНУ «Научный центр неврологии»
Russian Federation
Russian Federation
References
1. Пчелина С.Н. Альфа-синуклеин как биомаркер болезни Паркинсона // Анналы клинической и экспериментальной неврологии. 2011. Т. 5. № 4. С. 46-51.
2. Соболев В.Б., Худоерков Р.М. Иммуногистохимическое выявление α-синуклеина в слюнной железе как биомаркер болезни Паркинсона // Бюллетень Национального общества по изучению болезни Паркинсона и расстройств движений. 2017. № 2. С. 16-23.
3. Anwar S., Peters O., Millership S., Ninkina N., Doig N., Connor-Robson N., Threlfell S., Kooner G., Deacon R.M., Bannerman D.M., Bolam J.P., Chandra S.S., Cragg S.J., Wade-Martins R., Buchman V.L. Functional alterations to the nigrostriatal system in mice lacking all three members of the synuclein family // J. of Neurosci. 2011. No. 31(20). Pp. 7264-7274.
4. Arawaka S., Sato H., Sasaki A., Koyama S., Kato T. Mechanisms underlying extensive Ser129-phosphorylation in alpha-synuclein aggregates // Acta Neuropathologica Communications. 2017. No. 1(5). P. 48.
5. Braak H., Del Tredici K., Rüb U., de Vos R.A., Jansen Steur E.N., Braak E. Staging of brain pathology related to sporadic Parkinson's disease // Neurobiol. Aging. 2003. No. 24(2). Pp. 197-211.
6. Braak H., Sastre M., Bohl J.R., de Vos R.A., Del Tredici K. Parkinson's disease: lesions in dorsal horn layer I, involvement of parasympathetic and sympathetic pre- and postganglionic neurons // Acta Neuropathol. 2007. No. 113(4). Pp. 421-429.
7. Braz J.M., Nassar M.A., Wood J.N., Basbaum A.I. Parallel "pain" pathways arise from subpopulations of primary afferent nociceptor // Neuron. 2005. Vol. 47(6). Pp. 787-793.
8. Burre J., Sharma M., Sudhof T.C. Cell Biology and Pathophysiology of alpha-Synuclein // Cold Spring Harbor Perspectives in Medicine. 2017. pii: a024091. doi: 10.1101/cshperspect.a024091.
9. Del Tredici K., Braak H. Spinal cord lesions in sporadic Parkinson's disease // Acta Neuropathol. 2012. No. 124(5). Pp. 643-64.
10. Djelloul M., Holmqvist S., Boza-Serrano A., Azevedo C., Yeung M.S., Goldwurm S., Frisén J., Deierborg T., Roybon L. Alpha-Synuclein Expression in the Oligodendrocyte Lineage: an In Vitro and In Vivo Study Using Rodent and Human Models // Stem Cell Reports. 2015. No. 5(2). Pp. 174-84.
11. Donadio V., Incensi A., Rizzo G., Scaglione C., Capellari S., Fileccia E., Avoni P., Liguori R. Spine Topographical Distribution of Skin a-Synuclein Deposits in Idiopathic Parkinson Disease // J. of Neuropathology & Experimental Neurology. 2017. No. 5(76). Pp. 384-389.
12. Doppler K., Ebert S., Üçeyler N., Trenkwalder C., Ebentheuer J., Volkmann J., Sommer C. Cutaneous neuropathy in Parkinson’s disease: a window into brain pathology // Acta Neuropathol. 2014. No. 128(1). Pp. 99-109.
13. Giasson B.I., Duda J.E., Forman M.S., Lee V.M., Trojanowski J.Q. Prominent perikaryal expression of alpha- and beta-synuclein in neurons of dorsal root ganglion and in medullary neurons // Exp. Neurol. 2001. No. 172(2). Pp. 354-362.
14. Huang Z., Xu Z., Wu Y., Zhou Y. Determining nuclear localization of alpha-synuclein in mouse brains // Neuroscience. 2011. No. 199. Pp. 318-32.
15. Lin C.H., Chao C.C., Wu S.W., Hsieh P.C., Feng F.P., Lin Y.H., Chen Y.M., Wu R.M., Hsieh S.T. Pathophysiology of Small-Fiber Sensory System in Parkinson's Disease: Skin Innervation and Contact Heat Evoked Potential // Medicine (Baltimore). 2016. No. 95(10). E3058. doi: 10.1097/MD.0000000000003058.
16. Ma M.R., Hu Z.W., Zhao Y.F., Chen Y.X., Li Y.M. Phosphorylation induces distinct alpha-synuclein strain formation // Scientific Reports. 2016. No. 6. P. 37130. doi: 10.1038/srep37130.
17. Muntané G., Ferrer I., Martinez-Vicente M. α-synuclein phosphorylation and truncation are normal events in the adult human brain // Neuroscience. 2012. No. 200. Pp. 106-119.
18. Nam M.K., Han J.H., Jang J.Y., Yun S.E., Kim G.Y., Kang S., Rhim H. A novel link between the conformations, exposure of specific epitopes, and subcellular localization of alpha-synuclein // Biochimica et Biophysica Acta (BBA) - General Subjects. 2015. No. 12(1850). Pp. 2497-2505.
19. Oueslati A. Implication of Alpha-Synuclein Phosphorylation at S129 in Synucleinopathies: What Have We Learned in the Last Decade? // J. Parkinsons Dis. 2016. No. 6(1). Pp. 39-51.
20. Papachroni K., Ninkina N., Wanless J., Kalofoutis A.T., Gnuchev N.V., Buchman V.L. Peripheral sensory neurons survive in the absence of alpha- and gamma-synucleins // J. Mol. Neurosci. 2005. No. 25(2). Pp. 157-64.
21. Surguchov A. Intracellular Dynamics of Synucleins: "Here, There and Everywhere" // Int. Rev. Cell Mol. Biol. 2015. No. 320. Pp. 103-169.
22. Taguchi K., Watanabe Y., Tsujimura A., Tanaka M. Brain region-dependent differential expression of alpha-synuclein // J. Comp. Neurol. 2016. No. 524(6). Pp. 1236-58.
23. Tiunova A.A., Anokhin K.V., Saha A.R., Schmidt O., Hanger D.P., Anderton B.H., Davies A.M., Ninkina N.N., Buchman V.L. Chicken synucleins: cloning and expression in the developing embryo // Mech. Dev. 2000. No. 99(1-2). Pp. 195-198.
24. Vivacqua G., Casini A., Vaccaro R., Fornai F., Yu S., D'Este L. Different sub-cellular localization of alpha-synuclein in the C57BL6J mouse's central nervous system by two novel monoclonal antibodies // J. Chem. Neuroanat. 2011. No. 41(2). Pp. 97-110.
25. Pchelina S.N. Alfa-sinuklein kak biomarker bolezni Parkinsona [Alpha-synuclein as biomarker of Parkinson's disease]. Annaly klinicheskoj i ehksperimental'noj nevrologii [Annals of clinical and experimental neurology]. 2011. T. 5. No. 4. Pp. 46-51. (In Russian).
26. Sobolev V.B., Khudoerkov R.M. Immunogistohimicheskoe vyavlenie a-sinukleina v slyunnoj zheleze kak biomarker bolezni Parkinsona [Immunohistochemical detection of alpha-synuclein in salivary gland as biomarker of Parkinson disease]. Byulleten' Nacionalnogo obshchestva po izucheniyu bolezni Parkinsona i rasstrojstv dvizhenij [Bulletin of the National Society for the Study of Parkinson's Disease and Movement Disorders]. 2017. No. 2. Pp. 16-23. (In Russian).
27. Anwar S., Peters O., Millership S., Ninkina N., Doig N., Connor-Robson N., Threlfell S., Kooner G., Deacon R.M., Bannerman D.M., Bolam J.P., Chandra S.S., Cragg S.J., Wade-Martins R., Buchman V.L. Functional alterations to the nigrostriatal system in mice lacking all three members of the synuclein family. J. of Neurosci. 2011. No. 31(20). Pp. 7264-7274.
28. Arawaka S., Sato H., Sasaki A., Koyama S., Kato T. Mechanisms underlying extensive Ser129-phosphorylation in alpha-synuclein aggregates. Acta Neuropathologica Communications. 2017. No. 1(5). P. 48.
29. Braak H., Del Tredici K., Rüb U., de Vos R.A., Jansen Steur E.N., Braak E. Staging of brain pathology related to sporadic Parkinson's disease. Neurobiol. Aging. 2003. No. 24(2). Pp. 197-211.
30. Braak H., Sastre M., Bohl J.R., de Vos R.A., Del Tredici K. Parkinson's disease: lesions in dorsal horn layer I, involvement of parasympathetic and sympathetic pre- and postganglionic neurons. Acta Neuropathol. 2007. No. 113(4). Pp. 421-429.
31. Braz J.M., Nassar M.A., Wood J.N., Basbaum A.I. Parallel "pain" pathways arise from subpopulations of primary afferent nociceptor. Neuron. 2005. Vol. 47(6). Pp. 787-793.
32. Burre J., Sharma M., Sudhof T.C. Cell Biology and Pathophysiology of alpha-Synuclein. Cold Spring Harbor Perspectives in Medicine. 2017. pii: a024091. doi: 10.1101/cshperspect.a024091.
33. Del Tredici K., Braak H. Spinal cord lesions in sporadic Parkinson's disease. Acta Neuropathol. 2012. No. 124(5). Pp. 643-64.
34. Djelloul M., Holmqvist S., Boza-Serrano A., Azevedo C., Yeung M.S., Goldwurm S., Frisén J., Deierborg T., Roybon L. Alpha-Synuclein Expression in the Oligodendrocyte Lineage: an In Vitro and In Vivo Study Using Rodent and Human Models. Stem Cell Reports. 2015. No. 5(2). Pp. 174-84.
35. Donadio V., Incensi A., Rizzo G., Scaglione C., Capellari S., Fileccia E., Avoni P., Liguori R. Spine Topographical Distribution of Skin a-Synuclein Deposits in Idiopathic Parkinson Disease. J. of Neuropathology & Experimental Neurology. 2017. No. 5(76). Pp. 384-389.
36. Doppler K., Ebert S., Üçeyler N., Trenkwalder C., Ebentheuer J., Volkmann J., Sommer C. Cutaneous neuropathy in Parkinson’s disease: a window into brain pathology. Acta Neuropathol. 2014. No. 128(1). Pp. 99-109.
37. Giasson B.I., Duda J.E., Forman M.S., Lee V.M., Trojanowski J.Q. Prominent perikaryal expression of alpha- and beta-synuclein in neurons of dorsal root ganglion and in medullary neurons. Exp. Neurol. 2001. No. 172(2). Pp. 354-362.
38. Huang Z., Xu Z., Wu Y., Zhou Y. Determining nuclear localization of alpha-synuclein in mouse brains. Neuroscience. 2011. No. 199. Pp. 318-32.
39. Lin C.H., Chao C.C., Wu S.W., Hsieh P.C., Feng F.P., Lin Y.H., Chen Y.M., Wu R.M., Hsieh S.T. Pathophysiology of Small-Fiber Sensory System in Parkinson's Disease: Skin Innervation and Contact Heat Evoked Potential. Medicine (Baltimore). 2016. No. 95(10). E3058. doi: 10.1097/MD.0000000000003058.
40. Ma M.R., Hu Z.W., Zhao Y.F., Chen Y.X., Li Y.M. Phosphorylation induces distinct alpha-synuclein strain formation. Scientific Reports. 2016. No. 6. P. 37130. doi: 10.1038/srep37130.
41. Muntané G., Ferrer I., Martinez-Vicente M. α-synuclein phosphorylation and truncation are normal events in the adult human brain. Neuroscience. 2012. No. 200. Pp. 106-119.
42. Nam M.K., Han J.H., Jang J.Y., Yun S.E., Kim G.Y., Kang S., Rhim H. A novel link between the conformations, exposure of specific epitopes, and subcellular localization of alpha-synuclein. Biochimica et Biophysica Acta (BBA) - General Subjects. 2015. No. 12(1850). Pp. 2497-2505.
43. Oueslati A. Implication of Alpha-Synuclein Phosphorylation at S129 in Synucleinopathies: What Have We Learned in the Last Decade? J. Parkinsons Dis. 2016. No. 6(1). Pp. 39-51.
44. Papachroni K., Ninkina N., Wanless J., Kalofoutis A.T., Gnuchev N.V., Buchman V.L. Peripheral sensory neurons survive in the absence of alpha- and gamma-synucleins. J. Mol. Neurosci. 2005. No. 25(2). Pp. 157-64.
45. Surguchov A. Intracellular Dynamics of Synucleins: "Here, There and Everywhere". Int. Rev. Cell Mol. Biol. 2015. No. 320. Pp. 103-169.
46. Taguchi K., Watanabe Y., Tsujimura A., Tanaka M. Brain region-dependent differential expression of alpha-synuclein. J. Comp. Neurol. 2016. No. 524(6). Pp. 1236-58.
47. Tiunova A.A., Anokhin K.V., Saha A.R., Schmidt O., Hanger D.P., Anderton B.H., Davies A.M., Ninkina N.N., Buchman V.L. Chicken synucleins: cloning and expression in the developing embryo. Mech. Dev. 2000. No. 99(1-2). Pp. 195-198.
48. Vivacqua G., Casini A., Vaccaro R., Fornai F., Yu S., D'Este L. Different sub-cellular localization of alpha-synuclein in the C57BL6J mouse's central nervous system by two novel monoclonal antibodies. J. Chem. Neuroanat. 2011. No. 41(2). Pp. 97-110.
Review
For citations:
Voronkov D.N., Salnikova O.V., Khudoerkov R.M. Localization of total and Ser 129 phosphorylated α-synuclein in the rat peripheral nervous system (testing of different antibodies). Journal Biomed. 2018;(2):22-32. (In Russ.)
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