JoVE Journal > Medicine
Summary
Abstract
Introduction
Protocol
Results
Discussion
Disclosures
Acknowledgements
Materials
References
Automatic Translation
Medicine

Gene-miljø Interaktion Modeller til unmask modtagelighed mekanismer i Parkinsons sygdom

Published: January 07, 2014
doi:
10.3791/50960
, , ,
1Center for Health Sciences,SRI International, 2Department of Chemistry and Biochemistry,University of California-Santa Cruz

Summary

Lipoxygenase (LOX) isozymer kan generere produkter, der kan øge eller mindske neuroinflammation og neurodegeneration. En interaktionsundersøgelse mellem gen og miljø kunne identificere LOX isozymspecifikke virkninger. Ved hjælp af 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP) model af nigrostriatal skader i to LOX isozym-mangelfulde transgene linjer giver mulighed for sammenligning af bidraget fra LOX isozymer på dopaminergisk integritet og inflammation.

Abstract

Lipoxygenase (LOX) aktivitet har været impliceret i neurodegenerative lidelser såsom Alzheimers sygdom, men dens virkninger i Parkinsons sygdom (PD) patogenese er mindre forstået. Gen-miljø interaktion modeller har nytte i afmaskering virkningen af specifikke cellulære veje i toksicitet, som ikke kan observeres ved hjælp af en udelukkende genetisk eller toksikant sygdom model alene. For at vurdere, om forskellige LOX-isozymer selektivt bidrager til PD-relateret neurodegeneration, kan transgene (dvs. 5-LOX og 12/15-LOX mangelfulde) mus udfordres med et toksin, der efterligner celleskade og død i lidelsen. Her beskriver vi brugen af et neurotoksin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP), som producerer en nigrostriatal læsion for at belyse de forskellige bidrag fra LOX-isozymer til neurodegeneration relateret til PD. Brugen af MPTP i mus, og nonhuman primat, er veletableret til at opsummere nigrostriatal skader i PD. Omfanget af MPTP-induceret læsionering måles ved HPLC-analyse af dopamin og dets metabolitter og semi-kvantitativ western blot-analyse af striatum for tyrosinhydrafylase (TH), det hastighedsbegrænsende enzym til syntese af dopamin. For at vurdere inflammatoriske markører, som kan påvise LOX isozym-selektiv følsomhed, udføres glial fibrillary acidic protein (GFAP) og Iba-1 immunohistochemistry på hjernesektioner, der indeholder substantia nigra, og GFAP Western blot analyse udføres på striatal homogenater. Denne eksperimentelle tilgang kan give ny indsigt i gen-miljø interaktioner underliggende nigrostriatal degeneration og PD.

Introduction

Brug af gen-miljø interaktion modeller giver en tilgang til at efterligne risikofaktorer, der sandsynligvis påvirker idiopatisk Parkinsons sygdom (PD) og giver mulighed for at skelne mekanistiske indsigter, der er usandsynligt, at blive belyst ved brug af en genetisk eller toksikant system alene1,2. Her illustrerer vi dette punkt og beskriver anvendelsen af 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP) musemodel af nigrostriatal degeneration3 for bedre at forstå selektiviteten af lipoxygenase (LOX) isozymaktivitet på neuroinflammation ogtoksicitet 4. Mens en rolle for LOX isozymer er blevet bredt evalueret i perifere lidelser5,6 samt CNS sygdom, herunder slagtilfælde7 og Alzheimers sygdom8,9, den rolle, som familien af isozymer i nigrostriatal funktion og degeneration relateret til PD er ikke godt forstået og berettiger undersøgelse. MPTP neurotoksin demonstrerer præference degeneration af nigrostriatal vej og opsummerer striatal dopamin udtømning og nigral dopaminergiske celle tab, der ligger til grund motoriske funktionsnedsættelser i PD patienter10. Selv om denne model ikke gengiver den fulde cadre af nonmotoriske og motor pd adfærd og ærlig α-synuclein-positive Lewy krop patologi, det har været nyttigt at belyse nye mekanistiske mål, der bidrager til nigrostriatal skade og til tidlig oversættelsestest, da det er den bedst karakteriserede ikke-invasive model til rådighed til pålideligt at producere nigral celledød ledsaget af striatal dopamintab11-15. Bred brug af MPTP-musen, med paradigmer lige fra akut, subakut til kronisk16-18, har gjort det muligt for standardisering af dosering at resultere i mild til alvorlig nigrostriatal skade19,20 med aktivering af forskellige toksicitetsmekanismer afhængigt af behandlingsregimet18,21,22. Dette gør det derfor muligt at målrette et »læsionsvindue«, der kan resultere i øget eller reduceret nigrostriatal skade afhængigt af det terapeutiske middel eller den transgene model, der anvendes23-25.

Også afgørende for translationelle og opdagelse biologi undersøgelser er de teknikker, der anvendes til at vurdere skader og den dokumentation, sådanne metoder giver. For MPTP-musemodellen er etablerede målinger til vurdering af læsionering måling af markører for striatal dopaminergisk tone, herunder dopamin og dets metabolitter af HPLC, og vestlig blotanalyse af tyrosinhydroxylase (TH), det hastighedsbegrænsende enzym i dopaminsyntese og indikatorer for degenerative hændelser såsom glialaktivering ved hjælp af vestlig blot western analyse og immunohistochemistry4. Selv om disse er klassiske neurokemiske, biokemiske og histologiske procedurer, giver teknikkerne kritiske og reproducerbare udlæsninger om omfanget af skader inden for den nigrostriatale dopaminergiske vej, indikerer toksicitetsmekanismer og har vist sig at være værdifulde værktøjer til at forstå degenerative begivenheder i PD.

Protocol

Bemærk: Alle dyreforsøg og dyreplejemetoder bør godkendes af institutionens institutionelle dyrepleje- og brugsudvalg (IACUC). Den her beskrevne undersøgelse blev udført i overensstemmelse med retningslinjerne fra SRI Internationals IACUC. 1. Erhvervelse og vedligeholdelse af LOX-mangelfulde mus Køb 5-LOX-mangelfulde eller 12/15-LOX-mangelfulde mus og respektive stamme- og sexmatchede kontroller i en alder af 7-8 uger og g…

Representative Results

Dette toksineksponeringsparadigme kan producere en betydelig og påviselig 20% striatal dopaminudtømning hos MPTP- vs. saltvandsindsprøjtede dyr. Det er vigtigt at bemærke, at forskellige masser af MPTP kan give lidt mere eller mindre læsion; For bedre præcision anbefales således et indledende forsøg med wildtypemus inden brug i transgenik, når en ny masse neurotoksin udnyttes. Brugen af mild til moderat læsion gør det muligt at observere transgenets virkning; en alvorlig læsion kan producere en ‘gulveffekt’ …

Discussion

Udformningen af denne interaktionsundersøgelse om genmiljø gav os mulighed for at få nye oplysninger om 5-LOX isozymets dobbelte karakter i nigrostriatalvejen. Ved at udføre HPLC til måling af striatale monoaminer efter saltvands- eller MPTP-behandling i transgenik, der mangler 5-LOX-isozymet og deres littermates af vildtype, kunne vi konstatere, at dens mangel synes at være beskyttende under toksiske forhold (figur 1), men under normale forhold reducerer manglen på enzymet striatal dopaminniveau…

Disclosures

The authors have nothing to disclose.

Acknowledgements

Dette arbejde blev finansieret af National Institutes of Health NIGMS 056062.

Materials

1-Methyl-4-phenyl-1,2,3,6-tetra-hydropyridine hydrochloride (MPTP-HCL) Sigma-Aldrich M0896 for PD modeling
4% Formaldehyde (paraformaldehyde) solution, phosphate-buffered (PFA) American MasterTech Scientific BUP0157 for immersion fixation
Perchloric acid ACS reagent, 70% (PCA) Sigma-Aldrich 244252 for HPLC acid extraction
Tris Base Sigma-Aldrich T1503 for tissue homogenization
Ethylenediaminotetraacetic acid disodium salt dihydrate (EDTA) Sigma-Aldrich E1644 for tissue homogenization
Protease inhibitor cocktail Sigma-Aldrich P8340 for tissue homogenization
Phosphatase inhibitor cocktail Sigma-Aldrich P5726 for tissue homogenization
Sodium Hydroxide (NaOH) Sigma-Aldrich S5881 for Lowry protein assay
Sucrose, molecular biology, ≥99.5% (GC)  Sigma-Aldrich S0389 for cryoprotection
Phosphate buffered saline, powder, pH 7.4 (for 0.01 M PBS) Sigma-Aldrich P3813 for IHC
BCA Protein Assay Kit Pierce/Thermo 23225 for protein determination
Novex 12% Tris-Glycine Mini Gels 1.0 mm, 12-well Invitrogen/Life Technologies EC60052BOX for SDS-PAGE
NuPAGE LDS Sample Buffer (4x) Invitrogen/Life Technologies NP0007 for SDS-PAGE
Novex Sharp Prestained Protein Standard  Invitrogen/Life Technologies LC5800 protein ladder
Glycine Sigma-Aldrich G7126 for SDS-PAGE
Sodium dodecyl sulfate, electrophoresis, 98.5% (SDS) Sigma-Aldrich L3771 for SDS-PAGE
Methyl Alcohol, Anhydrous, Reagent  American MasterTech Scientific SPM1057C methanol for transfer
Sodium chloride (NaCl), ACS reagent Sigma-Aldrich S9888 saline and buffers
Nonfat dry milk powder Carnation n/a for immunoblotting
Ponceau S solution in 5% acetic acid  Sigma-Aldrich P7170 for immunoblotting
Anti-Tyrosine Hydroxylase (TH), sheep polyclonal Chemicon/Millipore AB1542 for immunofluorescence 
Anti-Tyrosine Hydroxylase (TH), rabbit polyclonal Pel-Freez Biologicals P40101-0 for immunoblotting
Anti-β Actin, rabbit Sigma-Aldrich A2066 for immunoblotting
Anti-Glial Fibrillary Acidic Protein (GFAP), rabbit polyclonal Chemicon/Millipore AB5804 for immunofluorescence
Anti-Glial Fibrillary Acidic Protein (GFAP), mouse monoclonal Covance Inc. SMI-22R for immunoblotting
Tween-20 Sigma-Aldrich P1379 for immunoblotting
Goat Anti-Rabbit IgG (H+L), Peroxidase Conjugated  Fisher Scientific 31462 for immunofluorescence
goat anti-sheep, peroxidase conjugated Pierce/Thermo 31480 for immunofluorescence
goat anti-mouse, peroxidase conjugated Pierce/Thermo 31430 for immunofluorescence
SuperSignal West Pico Chemiluminescent Substrate Pierce/Thermo 34078 for immunoblotting
CL-XPosure Film 7 in x 9.5 in  Pierce/Thermo 34089 for immunoblotting
Restore Western Blot Stripping Buffer  Pierce/Thermo 21059 for immunoblotting
Citric acid monohydrate, ACS reagent, ≥99.0%  Sigma-Aldrich C1909 for IHC
Normal Donkey Serum Millipore S30-100ML for IHC
Polyvinylpyrrolidone (PVP) Sigma-Aldrich P5288 for IHC
Bovine Serum Albumin (BSA), lyophilized Sigma-Aldrich A3294 for IHC
Triton X-100 Fisher Scientific BP151-01 for IHC
Donkey anti-Rabbit IgG, Alexa Fluor 568-labeled  Invitrogen/Life Technologies A10042 for IHC
Donkey Anti-Sheep IgG (H+L), FITC  Jackson ImmunoResearch 713-095-147 for IHC
VECTASHIELD Hard-Set Mounting Medium with DAPI Vector Laboratories H-1500 for IHC
Normal Goat Serum Millipore S26-100ML for IHC
VECTASTAIN ABC Kit (Rabbit IgG )  Vector Laboratories PK-4001 for IHC; 10 µl each of solutions A and B per 1 ml PBS (per instructions )
DAB Peroxidase Substrate Kit, 3,3’-diaminobenzidine Vector Laboratories SK-4100 for IHC; per 5 ml cold ddH2O, add 2 drops buffer stock solution, 2 drops DAB, and 1 drop H2O2 (H2O2 is added immediately before use)
Hydrogen peroxide, 30% Sigma-Aldrich 216763 for quench step in IHC
Rabbit anti-Iba1 Biocare Medicals CP290A for IHC
Cresyl Violet Solution, Regular Strength  FD Neurotechnologies PS102-01  counterstain for Iba1 IHC
95% Ethanol, reagent alcohol Sigma-Aldrich R8382 dehydration for IHC
100% Absolute ethanol Mallinckrodt  7019-10 dehydration for IHC
Acetic acid Sigma-Aldrich A6283 destaining for IHC
Xylene Sigma-Aldrich 534056 clearing agent for IHC
DPX Mountant Sigma-Aldrich 06522 mounting medium for DAB IHC
O.C.T. Compound – Frozen Section Embedding Medium  American MasterTech Scientific EMOCTCS embeddium medium for cryostat cutting
Potassium permanganate Sigma-Aldrich 223468 to decontaminate DAB solution
Dopamine hydrochloride Sigma-Aldrich H8502 for HPLC
3,4-Dihydroxyphenylacetic acid (DOPAC) Sigma-Aldrich 850217 for HPLC
Homovanillic acid (HVA) Sigma-Aldrich H1252 for HPLC
Perchloric acid (PCA) – 70% Sigma-Aldrich 244252 for HPLC
Sodium dihydrogen phosphate monohydrate Sigma-Aldrich 71504 for HPLC
Citric acid monohydrate Sigma-Aldrich C1909 for HPLC
1-Octanesulfonic acid sodium salt (OSA) Sigma-Aldrich O8380 for HPLC
EDTA Sigma-Aldrich E1644 for HPLC
Acetonitrile EMD AX0145-1 for HPLC
HPLC-grade distilled deionized water (ddH2O) Millipore for HPLC
0.22 µm GSTF membrane Millipore for filtration
Corning Netwells Sigma-Aldrich CLS3477 polystyrene insert with polyester mesh bottom, for IHC
[header]
Ultrasonic cell disrupter (Soniprep 150) MSE MSE.41371.274
Microcentrifuge Eppendorf 5414R
ESA MD-150 reverse-phase column  ESA
HPLC Pump (Ultimate 3000) Dionex ISO-3100BM
HPLC Autosampler (Ultimate 3000) Dionex WPS-3000TSL
Electrochemical detector ESA Coulochem III
Guard Cell ESA 5020
Analytical Cell ESA 5011A
Chromeleon software Dionex
Eclipse E400 Nikon E400 light/fluorescent microscope
Disposable mouse cage Ancare N10HT
Microfilter top Ancare N10MBT
[header]
5-LOX- deficient mice The Jackson Laboratory 004155
12/15-LOX-deficient mice The Jackson Laboratory 002778
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References

  1. Manning-Bog, A. B., Langston, J. W. Model fusion, the next phase in developing animal models for Parkinson's disease. Neurotox. Res. 11, 219-240 (2007).
  2. Vance, J. M., Ali, S., Bradley, W. G., Singer, C., Di Monte, D. A. Gene-environment interactions in Parkinson's disease and other forms of parkinsonism. Neurotoxicology. 31, 598-602 (2010).
  3. Heikkila, R. E., Hess, A., Duvoisin, R. C. Dopaminergic neurotoxicity of 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine in mice. Science. 224, 1451-1453 (1984).
  4. Chou, V. P., Holman, T. R., Manning-Bog, A. B. Differential contribution of lipoxygenase isozymes to nigrostriatal vulnerability. Neuroscience. 228, 73-82 (2013).
  5. Deschamps, J. D., Kenyon, V. A., Holman, T. R. Baicalein is a potent in vitro inhibitor against both reticulocyte 15-human and platelet 12-human lipoxygenases. Bioorg. Med.Chem. 14, 4295-4301 (2006).
  6. Weaver, J. R., et al. Integration of pro-inflammatory cytokines, 12-lipoxygenase and NOX-1 in pancreatic islet beta cell dysfunction. Mol. Cell Endocrinol. 358, 88-95 (2012).
  7. Yigitkanli, K., et al. Inhibition of 12/15-lipoxygenase as therapeutic strategy to treat stroke. Ann. Neurol. 73, 129-135 (2013).
  8. van Leyen, K., et al. Novel lipoxygenase inhibitors as neuroprotective reagents. J Neurosci. Res. 86, 904-909 (2008).
  9. Chu, J., Pratico, D. 5-lipoxygenase as an endogenous modulator of amyloid beta formation in vivo. Ann. Neurol. 69, 34-46 (2011).
  10. Langston, J. W., Ballard, P., Tetrud, J. W., Irwin, I. Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis. Science. 219, 979-980 (1983).
  11. Bove, J., Perier, C. Neurotoxin-based models of Parkinson's disease. Neuroscience. 211, 51-76 (2012).
  12. Beal, M. F. Neuroprotective effects of creatine. Amino Acids. 40, 1305-1313 (2011).
  13. Jackson-Lewis, V., Blesa, J., Przedborski, S. Animal models of Parkinson's disease. Parkinsonism Relat. Disord. 18, 183-185 (2012).
  14. Dauer, W., Przedborski, S. Parkinson's disease: mechanisms and models. Neuron. 39, 889-909 (2003).
  15. Wang, H., Shimoji, M., Yu, S. W., Dawson, T. M., Dawson, V. L. Apoptosis inducing factor and PARP-mediated injury in the MPTP mouse model of Parkinson's disease. Ann. N.Y. Acad. Sci. 991, 132-139 (2003).
  16. Petroske, E., Meredith, G. E., Callen, S., Totterdell, S., Lau, Y. S. Mouse model of Parkinsonism: a comparison between subacute MPTP and chronic MPTP/probenecid treatment. Neuroscience. 106, 589-601 (2001).
  17. Przedborski, S., et al. The parkinsonian toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP): a technical review of its utility and safety. J. Neurochem. 76, 1265-1274 (2001).
  18. Thomas, B., et al. Mitochondrial permeability transition pore component cyclophilin D distinguishes nigrostriatal dopaminergic death paradigms in the MPTP mouse model of Parkinson's disease. Antioxid. Redox. Signal. 16, 855-868 (2012).
  19. Sonsalla, P. K., Heikkila, R. E. The influence of dose and dosing interval on MPTP-induced dopaminergic neurotoxicity in mice. Eur. J. Pharmacol. 129, 339-345 (1986).
  20. Di Monte, D. A., et al. Relationship among nigrostriatal denervation, parkinsonism, and dyskinesias in the MPTP primate model. Mov. Disord. 15, 459-466 (2000).
  21. Lee, K. W., et al. Apoptosis signal-regulating kinase 1 mediates MPTP toxicity and regulates glial activation. PLoS One. 7, (2012).
  22. Jackson-Lewis, V., Przedborski, S. Protocol for the MPTP mouse model of Parkinson's disease. Nat. Protoc. 2, 141-151 (2007).
  23. Bolin, L. M., Strycharska-Orczyk, I., Murray, R., Langston, J. W., Di Monte, D. Increased vulnerability of dopaminergic neurons in MPTP-lesioned interleukin-6 deficient mice. J. Neurochem. 83, 167-175 (2002).
  24. Manning-Bog, A. B., et al. Increased vulnerability of nigrostriatal terminals in DJ-1-deficient mice is mediated by the dopamine transporter. Neurobiol. Dis. 27, 141-150 (2007).
  25. Quik, M., Di Monte, D. A. Nicotine administration reduces striatal MPP+ levels in mice. Brain Res. 917, 219-224 (2001).
  26. Markey, S. P., Johannessen, J. N., Chiueh, C. C., Burns, R. S., Herkenham, M. A. Intraneuronal generation of a pyridinium metabolite may cause drug-induced parkinsonism. Nature. 311, 464-467 (1984).
  27. Heikkila, R. E., Manzino, L., Cabbat, F. S., Duvoisin, R. C. Protection against the dopaminergic neurotoxicity of 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine by monoamine oxidase inhibitors. Nature. 311, 467-469 (1984).
  28. Crampton, J. M., Runice, C. E., Doyle, T. J., Lau, Y. S., Wilson, J. A. MPTP in mice: treatment, distribution and possible source of contamination. Life Sci. 42, 73-78 (1988).
  29. Yang, S. C., Markey, S. P., Bankiewicz, K. S., London, W. T., Lunn, G. Recommended safe practices for using the neurotoxin MPTP in animal experiments. Lab. Anim. Sci. 38, 563-567 (1988).
  30. Lau, Y. S., Novikova, L., Roels, C. MPTP treatment in mice does not transmit and cause Parkinsonian neurotoxicity in non-treated cagemates through close contact. Neuroscience research. 52, 371-378 (2005).
  31. Satoh, N., et al. Central hypothermic effects of some analogues of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and 1-methyl-4-phenylpyridinium ion (MPP). Neurosci. Lett. 80, 100-105 (1987).
  32. Fernagut, P. O., et al. Behavioral and histopathological consequences of paraquat intoxication in mice: effects of alpha-synuclein over-expression. Synapse. 61, 991-1001 (2007).
  33. Manning-Bog, A. B., McCormack, A. L., Purisai, M. G., Bolin, L. M., Di Monte, D. A. Alpha-synuclein overexpression protects against paraquat-induced neurodegeneration. J. Neurosci. 23, 3095-3099 (2003).
  34. Richfield, E. K., et al. Behavioral and neurochemical effects of wild-type and mutated human alpha-synuclein in transgenic mice. Exp. Neurol. 175, 35-48 (2002).
  35. Thomas, B., et al. Resistance to MPTP-neurotoxicity in alpha-synuclein knockout mice is complemented by human alpha-synuclein and associated with increased beta-synuclein and Akt activation. PloS one. 6, (2011).
  36. Smeyne, M., Goloubeva, O., Smeyne, R. J. Strain-dependent susceptibility to MPTP and MPP(+)-induced parkinsonism is determined by glia. Glia. 34, 73-80 (2001).
  37. Hamre, K., Tharp, R., Poon, K., Xiong, X., Smeyne, R. J. Differential strain susceptibility following 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration acts in an autosomal dominant fashion: quantitative analysis in seven strains of Mus musculus. Brain Res. 828, 91-103 (1999).
  38. Sedelis, M., et al. MPTP susceptibility in the mouse: behavioral, neurochemical, and histological analysis of gender and strain differences. Behav. Genet. 30, 171-182 (2000).
  39. Boyd, J. D., et al. Response to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) differs in mouse strains and reveals a divergence in JNK signaling and COX-2 induction prior to loss of neurons in the substantia nigra pars compacta. Brain Res. 1175, 107-116 (2007).
  40. Ookubo, M., Yokoyama, H., Kato, H., Araki, T. Gender differences on MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) neurotoxicity in C57BL/6 mice. Molecular and cellular endocrinology. 311, 62-68 (2009).
  41. Kenchappa, R. S., Diwakar, L., Annepu, J., Ravindranath, V. Estrogen and neuroprotection: higher constitutive expression of glutaredoxin in female mice offers protection against MPTP-mediated neurodegeneration. FASEB J. 18, 1102-1104 (2004).
  42. Jackson-Lewis, V., Jakowec, M., Burke, R. E., Przedborski, S. Time course and morphology of dopaminergic neuronal death caused by the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Neurodegeneration. 4, 257-269 (1995).
  43. Mizuno, Y., Sone, N., Saitoh, T. Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and 1-methyl-4-phenylpyridinium ion on activities of the enzymes in the electron transport system in mouse brain. J. Neurochem. 48, 1787-1793 (1987).
  44. Nicklas, W. J., Youngster, S. K., Kindt, M. V., Heikkila, R. E. M. P. T. P. MPP+ and mitochondrial function. Life Sci. 40, 721-729 (1987).
  45. Nicklas, W. J., Vyas, I., Heikkila, R. E. Inhibition of NADH-linked oxidation in brain mitochondria by 1-methyl-4-phenyl-pyridine, a metabolite of the neurotoxin, 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine. Life Sci. 36, 2503-2508 (1985).
  46. Wu, D. C., et al. Blockade of microglial activation is neuroprotective in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson disease. J. Neurosci. 22, 1763-1771 (2002).
  47. Kurkowska-Jastrzebska, I., Wronska, A., Kohutnicka, M., Czlonkowski, A., Czlonkowska, A. The inflammatory reaction following 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine intoxication in mouse. Exp. Neurol. 156, 50-61 (1999).
  48. Tatton, N. A., Kish, S. J. In situ detection of apoptotic nuclei in the substantia nigra compacta of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining. Neuroscience. 77, 1037-1048 (1997).
  49. Furuya, T., et al. Caspase-11 mediates inflammatory dopaminergic cell death in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson's disease. J Neurosci. 24, 1865-1872 (2004).
  50. Anderson, D. W., Bradbury, K. A., Schneider, J. S. Neuroprotection in Parkinson models varies with toxin administration protocol. Eur. J. Neurosci. 24, 3174-3182 (2006).

Tags

Keywords: Gene-environment InteractionLipoxygenaseParkinson’s DiseaseMPTPNeurodegenerative DisordersNigrostriatal DegenerationDopamineTyrosine HydroxylaseInflammationGFAPIba-1
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Chou, V. P., Ko, N., Holman, T. R., Manning-Boğ, A. B. Gene-environment Interaction Models to Unmask Susceptibility Mechanisms in Parkinson’s Disease. J. Vis. Exp. (83), e50960, doi:10.3791/50960 (2014).
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