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Methamphetamine increases Prion Protein and induces dopamine-dependent expression of protease resistant PrPsc

M. Ferrucci, L. Ryskalin, F. Biagioni, S. Gambardella, C. L. Busceti, A. Falleni, G. Lazzeri, F. Fornai


The cellular prion protein (PrPc) is physiologically expressed within selective brain areas of mammals. Alterations in the secondary structure of this protein lead to scrapie-like prion protein (PrPsc), which precipitates in the cell. PrPsc has been detected in infectious, inherited or sporadic neurodegenerative disorders. Prion protein metabolism is dependent on autophagy and ubiquitin proteasome. Despite not being fully elucidated, the physiological role of prion protein relates to chaperones which rescue cells under stressful conditions.
Methamphetamine (METH) is a widely abused drug which produces oxidative stress in various brain areas causing mitochondrial alterations and protein misfolding. These effects produce a compensatory increase of chaperones while clogging cell clearing pathways. In the present study, we explored whether METH administration modifies the amount of PrPc. Since high levels of PrPc when the clearing systems are clogged may lead to its misfolding into PrPsc, we further tested whether METH exposure triggers the appearance of PrPsc. We analysed the effects of METH and dopamine administration in PC12 and striatal cells by using SDS-PAGE Coomassie blue, immune- histochemistry and immune-gold electron microscopy. To analyze whether METH administration produces PrPsc aggregates we used antibodies directed against PrP following exposure to proteinase K or sarkosyl which digest folded PrPc but misfolded PrPsc. We fond that METH triggers PrPsc aggregates in DA-containing cells while METH is not effective in primary striatal neurons which do not produce DA. In the latter cells exogenous DA is needed to trigger PrPsc accumulation similarly to what happens in DA containing cells under the effects of METH. The present findings, while fostering novel molecular mechanisms involving prion proteins, indicate that, cell pathology similar to prion disorders can be mimicked via a DA-dependent mechanism by a drug of abuse.


autophagy; drugs of abuse; neurodegeneration; misfolded proteins; dopamine; striatum

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Aguib Y., Heiseke A., Gilch S., Riemer C., Baier M., Schätzl H.M., Ertmer A. Autophagy induction by trehalose counteracts cellular prion infection. Autophagy, 5: 361-369, 2009.

Battaglia G., Fornai, F., Busceti C.L., Aloisi G., Cerrito F., De Blasi A., Melchiorri D., Nicoletti F. Selective blockade of mGlu5 metabotropic glutamate receptors is protective against methamphetamine neurotoxicity. J. Neurosci., 22: 2135-2141, 2002.

Bolton D.C, McKinley M.P., Prusiner S.B. Identification of a protein that purifies with the scrapie prion. Science; 218: 1309-1311, 1982.

Castino R., Lazzeri G., Lenzi P., Bellio N., Follo C., Ferrucci M., Fornai F., Isidoro C. Suppression of autophagy precipitates neuronal cell death following low doses of methamphetamine. J. Neurochem., 106: 1426-1439, 2008.

Caughey B., Raymond G.J. Sulfated polyanion inhibition of scrapie-associated PrP accumulation in cultured cells. J Virol., 67: 643-650, 1993.

Chandra S., Fornai F., Kwon H.B., Yazdani U., Atasoy D., Liu X., Hammer R.E., Battaglia G., German D.C., Castello P.E., Südhof T.C. Double-knockout mice for alpha- and beta-synucleins: effect on synaptic functions. Proc. Natl. Acad. Sci. U S A., 101: 14966-14971, 2004.

Cherra S.J. 3rd, Dagda R.K., Chu C.T. Review: autophagy and neurodegeneration: survival at a cost? Neuropathol. Appl. Neurobiol., 36: 125-132, 2010.

Cohen-Kaplan V., Livneh I., Avni N., Cohen-Rosenzweig C., Ciechanover A. The ubiquitin-proteasome system and autophagy: Coordinated and independent activities. Int. J. Biochem. Cell. Biol., 79: 403-418, 2016

Conway K.A., Harper J.D., Lansbury P.T. Jr. Fibrils formed in vitro from.alpha-synuclein and two mutant forms linked to Parkinson's disease are typical.amyloid. Biochemistry, 39: 2552-2563, 2000.

Conway K.A., Rochet J.C., Bieganski R.M., Lansbury P.T. Jr. Kinetic stabilization of the alpha-synuclein protofibril by a dopamine-alpha-synuclein adduct. Science, 294: 1346-1349, 2001.

Cubells J.F, Rayport S., Rajendran G., Sulzer D. Methamphetamine neurotoxicity involves vacuolation of endocytic organelles and dopamine-dependent intracellular oxidative stress. J. Neurosci., 14: 2260-2271, 1994.

Das D., Luo X., Singh A., Gu Y., Ghosh S., Mukhopadhyay C.K., Chen S.G., Sy M.S., Kong Q., Singh N. Paradoxical role of prion protein aggregates in redox-iron induced toxicity. PLoS.One, 5: e11420, 2010.

Debeer, S.O.S., Baron T.G.M., Bencsik A.A. Immunohistochemistry of PrPsc Within Bovine Spongiform Encephalopathy Brain Samples with Graded Autolysis. J. Histochem. & Cytochem., 49: 1519-1524, 2001.

Debeer S.O.S, Baron T.G.M., Bencsik A.A. Transmissible Spongiform Encephalopathy Diagnosis Using PrPsc Immunohistochemistry on Fixed but Previously Frozen Brain Samples. J. Histochem. & Cytochem., 50: 611-616, 2002.

Deriziotis P., Tabrizi S.J. Prions and the proteasome. Biochim. Biophys. Acta., 1782: 713-722, 2008.

Desplats P., Lee H.J., Bae E.J., Patrick C., Rockenstein E., Crews L., Spencer B., Masliah E., Lee SJ. Inclusion formation and neuronal cell death through neuron-to-neuron transmission of alpha-synuclein. Proc. Natl. Acad. Sci. U S A., 106: 13010-13015, 2009.

Ferrucci M., Pasquali L., Paparelli A., Ruggieri S., Fornai F. Pathways of methamphetamine toxicity. Ann. N. Y. Acad. Sci., 1139: 177-85, 2008.

Fornai F., Giorgi F.S., Alessandrì M.G., Giusiani M., Corsini G.U. Effects of pretreatment with N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) on methamphetamine pharmacokinetics and striatal dopamine losses. J. Neurochem., 72: 777-784, 1999.

Fornai F., Lenzi P., Gesi M., Soldani P., Ferrucci M., Lazzeri G., Capobianco L., Battagli, G., De Blasi A., Nicoletti F., Paparelli A. Methamphetamine produces neuronal inclusions in the nigrostriatal system and in PC12 cells. J. Neurochem., 88: 114-123, 2004.

Fornai F., Ferrucci M., Gesi M., Bandettini di Poggio A., Giorgi F.S., Biagioni F., Paparelli A. A hypothesis on prion disorders: are infectious, inherited, and sporadic causes so distinct? Brain. Res. Bull., 69: 95-100, 2006.

Gilch S., Nunziante M., Ertmer A., Schätzl H.M. Strategies for eliminating PrP(c) as substrate for prion conversion and for enhancing PrP(Sc) degradation. Vet. Microbiol., 123: 377-386, 2007.

Haritani M, Spencer Y.I., Wells G.A. Hydrated autoclave pretreatment enhancement of prion protein immunoreactivity in formalin-fixed bovine spongiform encephalopathy-affected brain. Acta. Neuropathol. (Berl)., 87: 86–90, 1994.

Hegyi I., Hainfellner J.A, Flicker H., Ironside J., Hauw J.J., Tateishi J., Haltia M., Bugiani O., Aguzzi A., Budka H. Prion protein immunocytochemistry: reliable staining protocol, immunomorphology, and diagnostic pitfalls. Clinical Neuropathol., 16: 262-263, 1997.

Heiseke A., Aguib Y., Riemer C., Baier M., Schätzl H.M. Lithium induces clearance of protease resistant prion protein in prion-infected cells by induction of autophagy. J. Neurochem., 109: 25-34, 2009.

Heiseke A., Aguib Y., Schatzl H.M. Autophagy, prion infection and their mutual interactions. Curr. Issues. Mol. Biol., 12: 87-97, 2010.

Kovács G.G., Head M.W., Hegyi I., Bunn T.J., Flicker H., Hainfellner J.A, McCardle L., László L., Jarius, C., Ironside J.W., Budka H. Immunohistochemistry for the Prion Protein:Comparison of Different Monoclonal Antibodies in Human Prion Disease Subtypes. Brain. Pathol., 12: 1-11, 2002.

Kuczius T., Grassi J., Karch H., Groschup M.H. Binding of N- and C-terminal anti-prion protein antibodies generates distinct phenotypes of cellular prion proteins (PrPC) obtained from human, sheep, cattle and mouse. FEBS J., 274: 1492-1502, 2007.

Larsen K.E., Fon E.A., Hastings T.G., Edwards R.H., Sulzer D. Methamphetamine-induced degeneration of dopaminergic neurons involves autophagy and up regulation of dopamine synthesis. J. Neurosci., 22: 8951-8960, 2002.

Lazzeri G., Lenzi P., Gesi M., Ferrucci M., Fulceri F., Ruggieri S., Bruno V., Fornai F. In PC12 cells neurotoxicity induced by methamphetamine is related to proteasome inhibition. Ann N Y Acad Sci., 1074: 174-177 2006a.

Lazzeri G., Lenzi P., Ferrucci M., Fulceri F., Biagioni F., Blandini F., Paparelli A., Fornai F. Methamphetamine induces the expression of prion protein within striatum. NIDA Mini Conv. Abstr., 14, 2006b,

Lazzeri G., Lenzi P., Busceti C.L., Ferrucci M., Falleni A., Bruno V., Paparelli A., Fornai F. Mechanisms involved in the formation of dopamine-induced intracellular bodies within striatal neurons. J. Neurochem., 101: 1414-1427, 2007.

Lee S.J, Desplats P., Sigurdson C., Tsigelny I., Masliah E. Cell-to-cell transmission of non-prion protein aggregates. Nat. Rev. Neurol., 6: 702-706, 2010a.

Lee H.J., Suk J.E., Patrick C., Bae E.J., Cho J.H., Rho S., Hwang D., Masliah E., Lee S.J. Direct transfer of alpha-synuclein from neuron to astroglia causes inflammatory responses in synucleinopathies. J. Biol. Chem., 285: 9262-9272, 2010b.

Lenzi P., Lazzeri G., Biagioni F., Busceti C.L., Gambardella S., Salvetti A., Fornai F. The Autophagoproteasome a Novel Cell Clearing Organelle in Baseline and Stimulated Conditions. Front. Neuroanat., 10: 78, 2016.

Lezmi S., Bencsik A., Monks E., Petit T., Baron T. First case of feline spongiform encephalopathy in a captive cheetah born in France: PrPsc analysis in various tissues revealed unexpected targeting of kidney and adrenal gland. Histochem. Cell Biol., 119: 415-422, 2003.

Liautard J.P. Are prions misfolded molecular chaperones? FEBS. Lett., 294: 155-157, 1991.

Liautard J.P. Prions and molecular chaperones. Arch. Virol. Suppl., 7: 227-243, 1993.

Liautard J.P. Analytical background and discussion of the chaperone model of prion diseases. Acta. Biotheor., 47: 219-238, 1999.

Ma J., Lindquist S. De novo generation of a PrPSc-like conformation in living cells. Nat. Cell Biol., 1: 358-361, 1999.

Ma J., Lindquist. S. Conversion of PrP to a self-perpetuating PrPSc-like conformation in the cytosol. Science, 298: 1785-1788, 2002.

Milisav I., Šuput D., Ribarič S. Unfolded Protein Response and Macroautophagy in Alzheimer's, Parkinson's and Prion Diseases. Molecules, 20: 22718-22756, 2015.

Oh J.M., Shin H.Y., Park S.J., Kim B.H., Choi J.K., Choi E.K., Carp R.I., Kim Y.S. The involvement of cellular prion protein in the autophagy pathway in neuronal cells. Mol. Cell. Neurosci., 39: 238-247. 2008

Olanow C.W., Prusiner S.B. Is Parkinson’s disease a prion disorder? Proc. Natl. Acad. Sci. USA., 106: 12571-12572, 2009.

Pan K.M, Baldwin M., Nguyen J, Gasset M., Serban M, Groth D., Mehlhorn I., Huang Z., Fletterick R.J., Cohen F.E. Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc. Natl. Acad. Sci. USA., 90: 10962-10966, 1993.

Pasquali L., Lazzeri G., Isidoro C., Ruggieri S., Paparelli A., Fornai F. Role of autophagy during methamphetamine neurotoxicity. Ann. N. Y. Acad. Sci., 1139: 191-196, 2008.

Privat N., Sazdovitch V., Seilhean D., Laplanche J.L., Hauw J.J PrP. Immunohistochemistry: Different Protocols, Including a Procedure for Long Formalin Fixation, and a Proposed Schematic Classification for Deposits in Sporadic Creutzfeldt-Jakob Disease. Microsc. Res. Techn., 50: 26-31, 2000.

Prusiner S.B. Prions. Proc. Natl. Acad. Sci. USA., 95: 3363-13383, 1998.

Rose K., Goldberg M.P., Choi D.W. Cytotoxicity in murine neocortical cell cultures. pp. 46-60. In Tyson, C.A., Frazier J.M. (Eds), Methods in Toxicology San Diego Academics, San Diego, 1992.

Rosicarelli B., Serafini B., Sbriccoli M., Lu M., Cardone F., Pocchiari M., Aloisi, F. Migration of dendritic cells into the brain in a mouse model of prion disease. J. Neuroimmunol., 165: 114-120, 2005.

Schmidt C.J., Ritter J.K., Sonsalla P.K., Hanson G.R., Gibb J.W. Role of dopamine in the neurotoxic effects of methamphetamine. J. Pharmacol. Exp Ther., 233: 539-544, 1985.

Sigurdson C.J., Nilsson K.P.R., Hornemann S., Heikenwalder M., Manco G., Schwarz P., Ott D., Rülicke T., Liberski P.P., Julius C., Falsig J., Stitz L., Wüthrich K. & Aguzzi A. De novo generation of a transmissible spongiform encephalopathy by mouse transgenesis. Proc. Natl. Acad. Sci. USA., 106: 304-309, 2009.

Speldewinde S.H., Doronina V.A., Grant C.M. Autophagy protects against de novo formation of the [PSI+] prion in yeast. Mol. Biol. Cell. 26: 4541-4551, 2015.

Takahashi R.H., Tobiume M., Sato Y., Sata T., Gouras G.K., Takahashi H. Accumulation of cellular prion protein within dystrophic neuritis o amyloid plaques in the Alzheimer’s disease brain. Neuropathology, 31: 208-214, 2010.

Van Everbroeck B., Pals P., Martin J.J., Cras P. Antigen retrieval in prion protein immunohistochemistry. J. Histochem. Cytochem., 47: 1465-1470, 1999.

Zanusso G., Ferrari S., Cardone F., Zampieri P., Gelati M., Fiorini M., Farinazzo A., Gardiman M., Cavallaro T., Bentivoglio M., Righetti P.G., Pocchiari M., Rizzuto N., Monaco S. Detection of pathologic prion protein in the olfactory epithelium in sporadic Creutzfeldt-Jakob disease. N. Engl. J. Med., 348: 711-719, 2003.


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