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TTT 2015 Academic Conference Presentation

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TTT 2015 Academic Conference Presentation

  1. 1. Christine Yen [鄧思婷] Taiwan Tech Trek Academic Conference August 14, 2015 Modeling Neurodegenerative Diseases in Mice 1
  2. 2. Parkinson’s Disease (PD) 2 • Characteristics - Loss dopaminergic neurons in substantia nigra pars compacta (SNpc) • Causes: unknown, sporadic - Genetic, environmental factors • Prevalence - Primarily among elderly - 1-3% of population over 55 yrs http://what-when-how.com/wp-content/uploads/2012/04/tmp3675_thumb_thumb.jpg
  3. 3. Parkinson’s Disease • Pathophysiology -SNpc releases dopamine-> receptors in striatum - Dopamine deficiency - Striatum cannot signal motor thalamus, basal ganglia • Symptoms - Tremors, rigidity, bradykinesia difficulty walking - Dementia http://file.scirp.org/Html/2-2620020/24f49151-d68f-4540-bd71-c1b45e35562d.jpg http://discovermagazine.com/~/media/Images/Issues/2014/March/dopamine-chart.jpg Striatum SNpc 3
  4. 4. 6-OHDA Model 4 • What is 6-OHDA? ‒ Neurotoxin induces SNpc neurodegeneration • Dopaminergic selectivity - 6-OHDA structural analog: dopamine -Taken into SNpc via dopamine transporters (DAT) http://openi.nlm.nih.gov/imgs/512/250/2957224/2957224_PD2010-427810.002.png Before After
  5. 5. • Mechanism of neurotoxicity - Degradation by MAO - Generates H2O2, free radicals - Inhibits mitochondrial complex I • Why model PD? - No cure, limited treatments - Better understand pathology - Develop drugs: pre-clinical trials 6-OHDA Model of PD http://www.sciencedirect.com/science/article/pii/S030100820100003
  6. 6. Method of implementation • Stereotaxic surgery – Direct injections striatum – Unilateral vs bilateral injections • Retrograde degeneration of SNpc – Mimics PD: humans • Subjects – 9 mice injected w/ 3 mg/kg 6-OHDA – 3 controls injected with saline 6
  7. 7. Motor Outputs Left Brain Right Brain Controls left side of body Controls right side of body 6-OHDA lesion Motor impairments develop Motor function unaffected 7
  8. 8. Onset: PD Phenotypes • Successful model: replicates phenotypes - Symptoms typically emerge within weeks • Conduct behavioral assays - Assess degree motor deficits develop • Establish SOP’s: behavioral tests -Ensure protocols run smoothly http://www.nature.com/nrn/journal/v10/n7/fig_tab/nrn2652_F1.html8
  9. 9. Behavioral Assays: Overview Rotarod Test Adhesive Removal Test Forelimb Rearing Test 9
  10. 10. Rotarod Test • Assesses coordination, balancing abilities • Rod acceleration: 4-40 rpm • Walked 300s, rested: 15 min intervals • Record time spent continuously walking 10 (3 weeks post injection)
  11. 11. Rotarod in Action 11
  12. 12. Data Collection/Results 12
  13. 13. 13 Rotarod Performance: 6-OHDA groups vs control (training period) 180 190 200 210 220 230 240 250 260 270 280 7/29 7/30 7/31 6-OHDA groups Control group AverageTimewalked(s) Dates
  14. 14. 14 215 220 225 230 235 240 245 250 255 260 265 7/29 8/5 6-OHDA groups Control group Rotarod Performance: 6-OHDA groups vs control AverageTimewalked(s) Dates
  15. 15. Adhesive Removal Test • Measure sensorimotor deficits • Adhesives: alternately applied • Left, right forepaw • Measure time: sense, remove adhesives • Mice affected by 6-OHDA 15 Sticker
  16. 16. Forelimb Rearing Test • Assess forelimb motor impairment • Placed glass cylinder • Note forelimb favored: when rearing • Record frequency use: • Left, right or both forelimbs • Mice affected by 6-OHDA: • Ipsilateral forelimb dependence Forelimb Rearing Test 16
  17. 17. Modeling Alzheimer’s Disease (AD) 17
  18. 18. Morris Water Maze • Alzheimer’s disease behavioral assay • Tests spatial learning/memory • Developed SOP’s • Protocol runs smoothly • Establish appropriate environment 18
  19. 19. MWM Procedure ▪ Mice swim in tub: opaque water ▪ Find submerged platform ▪ Visual cues posted: walls ▪ Released: four different directions ▪ Factors of importance NW W SE SW 19
  20. 20. Conclusions ▪ Early stages: developing PD phenotypes ▪ SOP’s: identify, correct interfering factors ▪ Lack training ▪ Timing: trials ▪ Mouse behaviors 20
  21. 21. Acknowledgements ▪ My appreciation to:  Dr. Huang for her guidance  National Animal Laboratory Center, NARLabs  The mouse subjects ▪ Thank you all! 21

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