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The Effect of Centrophenoxine on Parkinson’s Disease . Victoria Wei. Need. Taken from Rajput AH, Offord KP, Beard CM, Kurland LT. Epidemiology of parkinsonism: incidence, classification, and mortality. Ann Neurol. 1984;16:278-282. .

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slide2
Need

Taken from Rajput AH, Offord KP, Beard CM, Kurland LT. Epidemiology of parkinsonism: incidence, classification, and mortality. Ann Neurol. 1984;16:278-282.

Figure 1 The amount of Parkinson’s disease cases per 100,000 people in the United States as age increases

knowledge base
Knowledge Base
  • Parkinson’s disease is a brain disorder involving the nerves.

Figure 2 The effects of Parkinson’s disease

http://www.spinstudios.co.uk/sa/pa3.jpg

knowledge base1
Knowledge Base

http://www.wormatlas.org/handbook/fig.s/IntroFIG6.jpg

Figure 3 The life cycle of C. elegans

knowledge base2
Knowledge Base

http://www.innovitaresearch.org/news/res/06042501_01.jpg

Figure 4 Lipofuscin in neurons of the human brain.

knowledge base3
Knowledge Base

http://commons.wikimedia.org/wiki/File:Centrophenoxine.svg

Figure 5 Molecular structure of centrophenoxine

literature review
Literature Review
  • Caldwin, et. al. (2008)

Figure 6

Figure 7

Both images taken from Caldwin, Guy A.; K.A. Caldwell. “Traversing a wormhole to Combat Parkinson’s disease.” Disease Models and Mechanisms. Volume 1. pp.000-000. 2008.

literature review1
Literature Review
  • Sutphin, et. al. (2009)

Figure 8

Auto fluorescent pigments present in Day 4 and Day 8 C. elegans

Sutphin, George; M. Kaeberlein. “Measuring Caenorhabditis elegans Life Span on Solid Media” JOVE. 2009.

literature review2
Literature Review
  • Gerstbrein, et. al. (2008)

Figure 9 Auto fluorescent pigments present in Day 7 and Day 12 C. elegans.

Gerstbrein, Beate; G. Stamatas; N. Kollias; M. Driscoll. “In vivspectrofluorimetry reveals endogenous biomarkers that report healthspan and dietary restriction in Caenorhabditis elegans.

literature review3
Literature Review
  • Application of centrophenoxine to the C. elegans decreased the rate of lipofuscin accumulation by an average of 41.3%. (Shulkin, et. al., 1978)

Figure 10 Table displaying effect of centrophenoxine on lipofuscin in C. elegans

Shulkin, D.J.; B.M. Zuckerman. “Spectrofluorometric analysis of the effect of centrophenoxine on lipofuscin accumulation in the nematode C. elegans.” Age. Volume 5. Pp. 50-53. 1982.

purpose
Purpose
  • The purpose of the experiment is to observe the effects of centrophenoxine on Parkinson’s disease in C. elegans

Hypothesis

  • Null- the symptoms of Parkinson’s disease will remain the same with or without the application of centrophenoxine.
  • Alternate- the symptoms of Parkinson’s disease will lessen with the application of centrophenoxine.
methodology

The Effects of Centrophenoxine on the development of Parkinson’s disease in C. elegans

Methodology

Wild type C. elegans and ham-1 (ot339) C. elegans obtained from the Caenorhabditis Genetics Center

N=80

Wild type C. elegans: N=40

ham-1(ot339) C. elegans: N=40

Given 6.8 mMcentrophenoxine

N=20

Control

N=20

Given 6.8 mMcentrophenoxine

N=20

Control

N=20

6.8 mMcentrophenoxine will be applied to the Nematode Growth Media for 21 days.

Use of 4\',6-diamidino-2-phenylindole (DAPI) to observe the amount of auto fluorescent pigment- lipofuscin- in C. elegans. GFP filters may also be used to observe whether the C. elegans DA neurons will be affected by the application of centrophenoxine

Statistical analysis

protocol
Protocol
  • C. elegans are grown in petri dishes containing Nematode Growth Media (NGM) from Carolina Biological and fed U.V. killed Escherichia coli.

Picture by author

Figure 11 Culturing the C. elegans in Petri dishes

protocol1
Protocol
  • Both Ampicillin and 5-Fluoro-2′-deoxyuridin will be used with NGM in the petri dishes with C. elegans

http://upload.wikimedia.org/wikipedia/commons/b/b6/Ampicillin_structure.svg

Figure 13 FUDR

Figure 12 Ampicillin

http://www.sigmaaldrich.com/structureimages/30/mfcd00006530.gif

E.coli + NGM with centrophenoxine+ Ampicillin + FUDR  using the DAPI and GFP filter to observe amount of lipofuscin and fluorescence present in both C. elegans groups

protocol2
Protocol

Sutphin, George; M. Kaeberlein. “Measuring Caenorhabditis elegans Life Span on Solid Media” JOVE. 2009.

Figure 14 Age synchronization of C. elegans

protocol3
Protocol

http://upload.wikimedia.org/wikipedia/commons/7/7a/DAPI.png

Figure 15 4\',6-diamidino-2-phenylindole (DAPI)

http://www.ion.ucl.ac.uk/images/Fluorescence-Microscope.jpg

http://www.bcgsc.ca/people/msleumer/htdocs/fact_sheet/image004.jpg

Figure 17 GFP expressed in C. elegans

Figure 16 Fluorescence microscope

do ability
Do-ability

Available for Purchase:

  • ham-1(ot339) and wild type C. elegans strains from CGC
  • DAPI and Sodium Azide from Sigma
  • NGM and OP50 E.coli from Carolina Biological
  • Centrophenoxine purchasable from Science Lab.com

Equipment already Acquired:

  • DAPI filter, GFP filter, fluorescent microscope
bibliography
Bibliography
  •  "About Parkinson Disease." National Parkinson Foundation. <”http://www.parkinson.org/Page.aspx?pid=225”>. 1996-2007.
  • Braungart, Evelyn; Gerlach, Manfred; Riederer; Peter, Baumeister, Ralf; and Hoener, Marius C. “Caenorhabditis elegans MPP+ Model of Parkinson’s Disease for High-throughout Drug Screening.” Neurodegenerative Disease. 2004. Volume 1: pgs 175-183.
  • Caldwin, Guy A.; K.A. Caldwell. “Traversing a wormhole to Combat Parkinson’s disease.” Disease Models and Mechanisms. Volume 1. pp.000-000. 2008.
  • Colleta, Susan. Introduction to C. elegans. Waksman Student Scholars. <http://avery.rutgers.edu/WSSP/StudentScholars/project/introduction/worms.html>. 2009
  • Gerstbrein, Beate; G. Stamatas; N. Kollias; M. Driscoll. “In vivspectrofluorimetry reveals endogenous biomarkers that report healthspan and dietary restriction in Caenorhabditis elegans.
  • Hall, D. H.; Z. F. Altun. “C. elegans Atlas.” Genetics Research,90 , pp 375-376. 2008.
  • Hunt, Sara S. The Aging Process. Washington D.C. April 2004.
  • Kenyon, Cynthia. “Environmental Factors and Gene Activities That Influence Life Span” C. elegans II. Cold Spring Harbor Press. 1997.
  • Kisiel, Marion J.; B. Zuckerman. “Effects of Centrophenoxine on the Nematode CaenorhabditisBriggsae” Age. Volume 1. Pp.17-20. January 1978.
  • Mc Naught, KS; P. Jenner. “Proteasomal function is impaired in substantianigra in Parkinson\'s disease “ Neuroscience Letters. Volume 297. pp. 191-194. 2001.
  • O\'Riordan ; A.M. Burnell. Intermediary metabolism in the dauer larva. II. The glyoxylate cycle and fatty acid oxidation. Comp. Biochem. Physiol. Volume 95. pp. 125-130. 1990.
  • Rajput AH, Offord KP, Beard CM, Kurland LT. Epidemiology of parkinsonism: incidence, classification, and mortality. Ann Neurol. 1984;16:278-282.
  • Schneider, Howard F.; C. Nandy. “Effects of Centrophenoxine on Lipofuscin Formation in Neuroblastoma Cells in Culture” Journal of Gerontology. Volume 32. Pp. 132-139. 1997.
  • Shulkin, D.J.; B.M. Zuckerman. “Spectrofluorometric analysis of the effect of centrophenoxine on lipofuscin accumulation in the nematode C. elegans.” Age. Volume 5. Pp. 50-53. 1982.
  • Sutphin, George; M. Kaeberlein. “Measuring Caenorhabditis elegans Life Span on Solid Media” JOVE. 2009.
  • “What is Parkinson’s?” American Parkinson Disease Association West Coast Office. <“http://www.apdawest.org/WhatIsParkinsons.html#2”>. 2009.
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