Pharynx of C. elegans http://www.wormbook.org/chapters/www_organformation/organformation.html
C. Elegans Life Cycle http://www.sfu.ca/biology/faculty/ hutter/hutterlab/research/pictures /Ce_life_cycle.gif
Fig. 2 Blueberry polyphenols extend lifespan and slow aging in C. elegans Literature Review Wilson et. al. 2006
Literature Review Figure 3: Mean lifespan of different genotypes of C. elegans
Literature Review Figure 4: Structural and functional declines in C. elegans pharynx from days 2-10. Chow et. al. 2006
Literature Review James Joseph (2007-2009) Tested and still testing fruit polyphenols and their effects on aging and the brain of mice Tested effects of blackberries, blueberries, strawberries, plum juice, and other berry fruits
Purpose Therefore the purpose of this experiment is to test the ability of fruit polyphenols in the prolonging of lifespan in C. elegans. Hypothesis Null Hypothesis: The fruit polyphenols will show no increase on the lifespan of C. elegans. Alternate Hypothesis: The fruit polyphenols will successfully increase the lifespan of C. elegans.
Methodology Caenorhabditis elegans Short lifespan of about 2-3 weeks, rapid generation time, and experiemental flexibilty Control Medium Sample Size = 5 Medium + Fed crude blueberry extract (Vaccinium angustifolium) Sample Size = 5 Measurement Average pumping rate of pharynx contraction is recorded with the use of Motic Plus Camera attached to a stereomicroscope. Statistical Analysis Medium + Fed crude blackberry extract Sample Size = 5 Medium + Fed crude strawberry extract Sample Size = 5 Medium + Fed crude Plum extract Sample Size = 5
Protocols for Measurement Commercially prepared single strength wild blueberry juice (Vaccinium angustifolium) is applied to a preconditioned C18 Column C18 column is washed with water to remove fructose, glucose, and organic acids of blueberries, then with 100% methanol to obtain the total polyphenolic fraction http://www.waters.com/waters/partDetail.htm?local e=101&partNumber=WAT066224
Protocols for Measurement Fruit polyphenol extracts is added to NGM agar plates of C. elegans on the first day of the lifespan assay Lifespan scoring is initiated after hermaphrodites complete their final larval molt, on the first day of adulthood Pharynx pumping rates are measured on days 4, 6, 8, and 10 of C. elegans adulthood
Protocols for Measurement Pharynx contraction rate recorded with Motic Plus camera attached to a stereomicroscope Pumps per minute measured http://www.technicalvideo.com/ photos/ElegansPharynx.jpg
Doability Motic Plus with Stereomicroscope available in lab for pharynx contraction observation Fruit polyphenols are easily obtainable C. elegans are easily obtainable and culturable
BudgetVendor Catalogue # PageItem Item # Quantit y Unit $$ Total $$ Carolina Science 2009 78248 C. elegans N2 (wild type) NP-17- 3500 1 $10.95 (per culture) $10.95 Caroline Science 2009 78248 Nematode Growth Agar NP-17- 3520 5 $6.25 (per 135 ml bottle) $31.25 https://www.wat ers.com/waters/ partDetail.htm?p artNumber=WA T066224 n/an/a Symmetry C18 Column, 3.5 µm, 4.6 x 75 mm WAT066 224 1$543.00$543.00 Carolina Science Websiten/an/aMethanol 8749501 $5.75 per 500 mL $5.75 Total Price: $590.95
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