Early Parkinson’s Disease-Like Pathology in a Transgenic Mouse Model Involves a Decreased Cst3 mRNA Expression But Not Neuroinflammatory Response in the Brain

T.A. Korolenko 1 , A.B. Shintyapina 3 , V.M. Belichenko 1 , A.B. Pupyshev 1 , A.A. Akopyan 1 , L.A. Fedoseeva 1 , 4 , G.S. Russkikh 5 , V.A. Vavilin 3 , M.V. Tenditnik 1 , C.-L. Lin 6 , T.G. Amstislavskaya 2 , and M.A. Tikhonova 1
  • 1 Laboratory of Experimental Models of Neurodegenerative Processes, Department of Experimental Neuroscience, Scientific Research Institute of Physiology and Basic Medicine, Russia, Novosibirsk
  • 2 Laboratory of Translational Biopsychiatry, Department of Experimental Neuroscience, Scientific Research Institute of Physiology and Basic Medicine, Russia, Novosibirsk
  • 3 Laboratory of Drug Metabolism and Pharmacokinetics, Scientific Research Institute of Molecular Biology and Biophysics, Federal Research Center for Basic and Translational Medicine, Russia, Novosibirsk
  • 4 Laboratory of Evolutionary Genetics, Federal Research Center “Institute of Cytology and Genetics”, Siberian Branch of the Russian Academy of Sciences, Russia, Novosibirsk
  • 5 Scientific Research Institute of Biochemistry, Federal Research Center for Basic and Translational Medicine, , Russia, Novosibirsk
  • 6 Institute of Medicine, Chung Shan Medical University, Taichung


Pathological aggregation and accumulation of α-synuclein in neurons play a core role in Parkinson’s disease (PD) while its overexpression is a common PD model. Autophagy-lysosomal pathways are general intraneural mechanisms of protein clearance. Earlier a suppressed autophagy in the brain of young transgenic mice overexpressing the А53Т-mutant human α-synuclein (mut(PD)) was revealed. Previous studies have recognized that Cystatin C displays protective activity against neurodegeneration. This cysteine protease inhibitor attracts particular attention as a potential target for PD treatment related to autophagy modulation. Here we evaluated the mRNA levels of Cst3 encoding Cystatin C in different brain structures of 5 m.o. mut(PD) mice at standard conditions and after the chronic treatment with a neuroprotective agent, ceftriaxone (100 mg/kg, 36 days). The inflammatory markers, namely, microglial activation by IBA1 expression and mRNA levels of two chitinases genes (Chit1, Chia1), were also assessed but no significant difference was found between control and transgenic mice. Cst3 mRNA levels were significantly reduced in the striatum and amygdala in the transgenic PD model. Furthermore, this was associated with autophagy decline and might be added to early signs of synucleinopathy development. We first demonstrated the modulation of mRNA levels of Cst3 and autophagy marker Becn1 in the brain by ceftriaxone treatment. Taken together, the results support the potential of autophagy modulation through Cystatin C at early stages of PD-like pathology.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • 1. Rocha EM, De Miranda B, Sanders LH. Alpha-synuclein: Pathology, mitochondrial dysfunction and neuroinfl ammation in Parkinson’s disease. Neurobiol Dis. 2018;109(Pt B):249–57. https://doi.org/10.1016/j.nbd.2017.04.004

  • 2. Jagmag SA, Tripathi N, Shukla SD, Maiti S, Khurana S. Evaluation of Models of Parkinson’s Disease. Front Neurosci. 2016:503. https://doi.org/10.3389/fnins.2015.00503

  • 3. Deleidi M, Maetzler W. Protein clearance mechanisms of alpha-synuclein and amyloid-Beta in lewy body disorders. Int J Alzheimers Dis. 2012;2012:391438. https://doi.org/10.1155/2012/391438

  • 4. Cerri S, Blandini F. Role of Autophagy in Parkinson’s Disease. Curr Med Chem. 2019;26(20):3702–18. https://doi.org/10.2174/0929867325666180226094351

  • 5. Seronie-Vivien S, Delanaye P, Pieroni L, Mariat C, Froissart M, Cristol JP, et al. Cystatin C: current position and future prospects. Clin Chem Lab Med. 2008;46(12):1664–86. https://doi.org/10.1515/CCLM.2008.336

  • 6. Amin F, Khan MS, Bano B. Mammalian cystatin and protagonists in brain diseases. J Biomol Struct Dyn. 2020;38(7):2171–96. https://doi.org/10.1080/07391102.2019.1620636

  • 7. Keppler D. Towards novel anti-cancer strategies based on cystatin function. Cancer Lett. 2006;235(2):159–76. https://doi.org/10.1016/j.canlet.2005.04.001

  • 8. Korolenko TA, Shintyapina AB, Pupyshev AB, Akopyan AA, Russkikh GS, Dikovskaya MA, et al. The regulatory role of cystatin C in autophagy and neurodegeneration. Vavilov Journal of Genetics and Breeding. 2019;23(4):390–7. https://doi.org/10.18699/vj19.507

  • 9. Pérez-González R, Sahoo S, Gauthier SA, Kim Y, Li M, Kumar A, et al. Neuroprotection mediated by cystatin C-loaded extracellular vesicles. Sci Rep. 2019;9(1):11104. https://doi.org/10.1038/s41598-019-47524-7

  • 10. Kaur G, Levy E. Cystatin C in Alzheimer’s disease. Front Mol Neurosci. 2012;5:79. https://doi.org/10.3389/fnmol.2012.00079

  • 11. Gauthier S, Kaur G, Mi W, Tizon B, Levy E. Protective mechanisms by cystatin C in neurodegenerative diseases. Front Biosci (Schol Ed). 2011;3:541–54.

  • 12. Hu WD, Chen J, Mao CJ, Feng P, Yang YP, Luo WF, et al. Elevated Cystatin C Levels Are Associated with Cognitive Impairment and Progression of Parkinson Disease. Cogn Behav Neurol. 2016;29(3):144–9. http://dx.doi.org/10.1097/wnn.0000000000000100

  • 13. Xu L, Sheng J, Tang Z, Wu X, Yu Y, Guo H, et al. Cystatin C prevents degeneration of rat nigral dopaminergic neurons: in vitro and in vivo studies. Neurobiol Dis. 2005;18(1):152–65. https://doi.org/10.1016/j.nbd.2004.08.012

  • 14. Zou J, Chen Z, Wei X, Chen Z, Fu Y, Yang X, et al. Cystatin C as a potential therapeutic mediator against Parkinson’s disease via VEGF-induced angiogenesis and enhanced neuronal autophagy in neurovascular units. Cell Death Dis. 2017;8(6):e2854. https://doi.org/10.1038/cddis.2017.240

  • 15. Tizon B, Sahoo S, Yu H, Gauthier S, Kumar AR, Mohan P, et al. Induction of autophagy by cystatin C: a mechanism that protects murine primary cortical neurons and neuronal cell lines. PLoS One. 2010;5(3):e9819. https://doi.org/10.1371/journal.pone.0009819

  • 16. Yimer EM, Hishe HZ, Tuem KB. Repurposing of the β-Lactam Antibiotic, Ceftriaxone for Neurological Disorders: A Review. Front Neurosci. 2019;13:236. https://doi.org/10.3389/fnins.2019.00236

  • 17. Tai CH, Bellesi M, Chen AC, Lin CL, Li HH, Lin PJ, et al. A new avenue for treating neuronal diseases: Ceftriaxone, an old antibiotic demonstrating behavioral neuronal effects. Behav Brain Res. 2019;364:149–56. https://doi.org/10.1016/j.bbr.2019.02.020

  • 18. Gelders G, Baekelandt V, Van der Perren A. Linking Neuroinfl ammation and Neurodegeneration in Parkinson’s Disease. J Immunol Res. 2018;2018:4784268. https://doi.org/10.1155/2018/4784268

  • 19. Li T, Le W. Biomarkers for Parkinson’s Disease: How Good Are They? Neurosci Bull. 2020;36(2):183–94. https://doi.org/10.1007/s12264-019-00433-1

  • 20. Tikhonova MA, Amstislavskaya TG, Belichenko VM, Fedoseeva LA, Kovalenko SP, Pisareva EE, et al. Modulation of the expression of genes related to the system of amyloid-beta metabolism in the brain as a novel mechanism of ceftriaxone neuroprotective properties. BMC Neurosci. 2018;19(Suppl 1):13. https://doi.org/10.1186/s12868-018-0412-5

  • 21. Tikhonova MA, Ho SC, Akopyan AA, Kolosova NG, Weng JC, Meng WY, et al. Neuroprotective effects of ceftriaxone treatment on cognitive and neuronal deficits in a rat model of accelerated senescence. Behav Brain Res. 2017;330:8–16. https://doi.org/10.1016/j.bbr.2017.05.002

  • 22. Iwai-Kanai E, Yuan H, Huang C, Sayen MR, Perry-Garza CN, Kim L, et al. A method to measure cardiac autophagic fl ux in vivo. Autophagy. 2008;4(3):322–9. https://doi.org/10.4161/auto.5603

  • 23. Ruijter JM, Ramakers C, Hoogaars WM, Karlen Y, Bakker O, van den Hoff MJ, et al. Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res. 2009;37(6):e45. https://doi.org/10.1093/nar/gkp045

  • 24. Pupyshev AB, Tikhonova MA, Akopyan AA, Tenditnik MV, Dubrovina NI, Korolenko TA. Therapeutic activation of autophagy by combined treatment with rapamycin and trehalose in a mouse MPTP-induced model of Parkinson’s disease. Pharmacol Biochem Behav. 2019;177:1–11. https://doi.org/10.1016/j.pbb.2018.12.005

  • 25. Streit WJ, Xue QS, Tischer J, Bechmann I. Microglial pathology. Acta Neuropathol Commun. 2014;2:142. https://doi.org/10.1186/s40478-014-0142-6

  • 26. Correale J, Fiol M. Chitinase effects on immune cell response in neuromyelitis optica and multiple sclerosis. Mult Scler. 2011;17(5):521–31. https://doi.org/10.1177/1352458510392619

  • 27. Hall S, Surova Y, Öhrfelt A; Swedish BioFINDER Study, Blennow K, Zetterberg H, Hansson O. Longitudinal Measurements of Cerebrospinal Fluid Biomarkers in Parkinson’s Disease. Mov Disord. 2016;31(6):898–905. https://doi.org/10.1002/mds.26578

  • 28. Xiao Q, Yu W, Tian Q, Fu X, Wang X, Gu M, et al. Chitinase1 contributed to a potential protection via microglia polarization and Aβ oligomer reduction in D-galactose and aluminum-induced rat model with cognitive impairments. Neuroscience. 2017;355:61–70. https://doi.org/10.1016/j.neuroscience.2017.04.050

  • 29. Steinacker P, Verde F, Fang L, Feneberg E, Oeckl P, Roeber S, et al. Chitotriosidase (CHIT1) is increased in microglia and macrophages in spinal cord of amyotrophic lateral sclerosis and cerebrospinal fl uid levels correlate with disease severity and progression. J Neurol Neurosurg Psychiatry. 2018;89(3):239–47. http://dx.doi.org/10.1136/jnnp-2017-317138

  • 30. Zhang W, Dallas S, Zhang D, Guo JP, Pang H, Wilson B, et al. Microglial PHOX and Mac-1 are essential to the enhanced dopaminergic neurodegeneration elicited by A30P and A53T mutant alpha-synuclein. Glia. 2007;55(11):1178–88. https://doi.org/10.1002/glia.20532

  • 31. Sirabella R, Sisalli MJ, Costa G, Omura K, Ianniello G, Pinna A, et al. NCX1 and NCX3 as potential factors contributing to neurodegeneration and neuroinfl ammation in the A53T transgenic mouse model of Parkinson’s Disease. Cell Death Dis. 2018;9(7):725. https://doi.org/10.1038/s41419-018-0775-7

  • 32. Gao HM, Zhang F, Zhou H, Kam W, Wilson B, Hong JS. Neuroinfl ammation and alpha-synuclein dysfunction potentiate each other, driving chronic progression of neurodegeneration in a mouse model of Parkinson’s disease. Environ Health Perspect. 2011;119(6):807–14. https://doi.org/10.1289/ehp.1003013

  • 33. Wu Q, Yang X, Zhang Y, Zhang L, Feng L. Chronic mild stress accelerates the progression of Parkinson’s disease in A53T α-synuclein transgenic mice. Exp Neurol. 2016;285(Pt A):61–71. https://doi.org/10.1016/j.expneurol.2016.09.004

  • 34. Pupyshev AB, Korolenko TA, Akopyan AA, Amstislavskaya TG, Tikhonova MA. Suppression of autophagy in the brain of transgenic mice with overexpression of А53Т-mutant α-synuclein as an early event at synucleinopathy progression. Neurosci Lett. 2018;672:140–4. https://doi.org/10.1016/j.neulet.2017.12.001

  • 35. Klionsky DJ, Abdalla FC, Abeliovich H, Abraham RT, Acevedo-Arozena A, Adeli K, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy. 2012;8(4):445–544. http://dx.doi.org/10.4161/auto.19496

  • 36. Ahumada-Castro U, Silva-Pavez E, Lovy A, Pardo E, Molgό J, Cárdenas C. MTOR-independent autophagy induced by interrupted endoplasmic reticulum-mitochondrial Ca2+ communication: a dead end in cancer cells. Autophagy. 2019;15(2):358–61. https://doi.org/10.1080/15548627.2018.1537769

  • 37. Williams A, Sarkar S, Cuddon P, Ttofi EK, Saiki S, Siddiqi FH, et al. Novel targets for Huntington’s disease in an mTOR-independent autophagy pathway. Nat Chem Biol. 2008;4(5):295–305. https://doi.org/10.1038/nchembio.79

  • 38. Fang Z, Feng Y, Li Y, Deng J, Nie H, Yang Q, et al. Neuro-protective Autophagic Flux Induced by Hyperbaric Oxygen Preconditioning is Mediated by Cystatin C. Neurosci Bull. 2019;35(2):336–46. https://doi.org/10.1007/s12264-018-0313-8

  • 39. Wang R, Chen Z, Fu Y, Wei X, Liao J, Liu X, et al. Plasma Cystatin C and High-Density Lipoprotein Are Important Biomarkers of Alzheimer’s Disease and Vascular Dementia: A Cross-Sectional Study. Front Aging Neurosci. 2017;9:26. https://doi.org/10.3389/fnagi.2017.00026

  • 40. Kaur G, Mohan P, Pawlik M, DeRosa S, Fajiculay J, Che S, et al. Cystatin C rescues degenerating neurons in a cystatin B-knockout mouse model of progressive myoclonus epilepsy. Am J Pathol. 2010;177(5):2256–67. https://doi.org/10.2353/aj-path.2010.100461

  • 41. Chen WW, Cheng X, Zhang X, Zhang QS, Sun HQ, Huang WJ, et al. The expression features of serum Cystatin C and homo-cysteine of Parkinson’s disease with mild cognitive dysfunction. Eur Rev Med Pharmacol Sci. 2015;19(16):2957–63.

  • 42. Xiong Y, Mahmood A, Chopp M. Current understanding of neuroinfl ammation after traumatic brain injury and cell-based therapeutic opportunities. Chin J Traumatol. 2018;21(3):137–51. https://doi.org/10.1016/j.cjtee.2018.02.003

  • 43. Ho SC, Hsu CC, Pawlak CR, Tikhonova MA, Lai TJ, Amstislavskaya TG, et al. Effects of ceftriaxone on the behavioral and neuronal changes in an MPTP-induced Parkinson’s disease rat model. Behav Brain Res. 2014;268:177–84. https://doi.org/10.1016/j.bbr.2014.04.022

  • 44. Weng JC, Tikhonova MA, Chen JH, Shen MS, Meng WY, Chang YT, et al. Ceftriaxone prevents the neurodegeneration and decreased neurogenesis seen in a Parkinson’s disease rat model: An immunohistochemical and MRI study. Behav Brain Res. 2016;305:126–39. https://doi.org/10.1016/j.bbr.2016.02.034

  • 45. Cui C, Cui Y, Gao J, Sun L, Wang Y, Wang K, et al. Neuro-protective effect of ceftriaxone in a rat model of traumatic brain injury. Neurol Sci. 2014;35(5):695–700. https://doi.org/10.1007/s10072-013-1585-4

  • 46. Huh C, Nagle JW, Kozak CA, Abrahamson M, Karlsson S. Structural organization, expression and chromosomal mapping of the mouse cystatin-C-encoding gene (Cst3). Gene. 1995;152(2):221–6. https://doi.org/10.1016/0378-1119(94)00728-B

  • 47. Abdel-Daim MM, El-Sayed YS, Eldaim MA, Ibrahim A. Nephro-protective efficacy of ceftriaxone against cisplatin-induced subchronic renal fibrosis in rats. Naunyn Schmiedebergs Arch Pharmacol. 2017;390:301–9. https://doi.org/10.1007/s00210-016-1332-5

  • 48. Sarafian TA, Littlejohn K, Yuan S, Fernandez C, Cilluffo M, Koo BK, et al. Stimulation of synaptoneurosome glutamate release by monomeric and fibrillated α synuclein. J Neurosci Res. 2017;95(9):1871–87. https://doi.org/10.1002/jnr.24024

  • 49. Lopez-Colome MA, Martinez-Lozada Z, Guillem AM, Lopez E, Ortega A. Glutamate transporter-dependent mTOR phosphorylation in Muller glia cells. ASN Neuro. 2012:4(5):e00095. http://dx.doi.org/10.1042/AN20120022


Journal + Issues