William Kim and Nicole Kiess. Abstract Sarcopenia is defined as the loss of muscle mass, strength,...
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Transcript of William Kim and Nicole Kiess. Abstract Sarcopenia is defined as the loss of muscle mass, strength,...
William Kim and Nicole Kiess
Abstract
Sarcopenia is defined as the loss of muscle mass, strength, and function associated with aging. As aging takes place, there is a sharp decrease in muscle mass and strength seen in both physically active individuals and physically inactive individuals. Sarcopenia is a major feature of aging in both humans and C. elegans, a model organism used for studying aging and longevity. Blueberry polyphenols have been found to increase the lifespan of C. elegans, seen in the pharynx contraction rate in adult C. elegans. C. elegans pumping rate of the pharynx is an indication of the age for the pumping rate of the pharynx decreases with age. Using various concentrations of different fruit polyphenols the lifespan of C. elegans will be measured by observing the pharynx contraction rate. Seventy-five worms will be fed two different concentrations of plum polyphenols, fifty C. elegans will be fed three different concentrations of strawberry polyphenols, and there will be one-hundred fifty worms in the control group. They all will be fed an equal amount of each fruit polyphenol. The average pumping rate of the pharynx per minute will be measured with the use of a Motic camera attached to a Zeiss Axiovert 40 CFL microscope.
The Effect of Fruit Polyphenols on the Longevity of C. elegans
Figure 1: Muscle Mass Associated with Aging
Need
Caenorhabditis elegans
http://www.imsc.res.in/~sitabhra/research/neural/celegans/celegans.gif
http://shirleywho.files.wordpress.com/2008/09/c-elegans_esa.jpg
Image 1: C. elegans external appearance Image 2: C. elegans internal appearance
C. Elegans Life Cycle
http://www.sfu.ca/biology/faculty/hutter/hutterlab/research/pictures/Ce_life_cycle.gif
Image 3: C. elegans Life Cycle
Pharynx of C. elegans
http://www.wormbook.org/chapters/www_organformation/organformation.html
Image 4: Pharyngeal Structure of Caenorhabditis elegans
Fig. 2 Blueberry polyphenols extend lifespan and slow aging in C. elegans
Literature Review
Wilson et. al. 2006
Literature Review
Figure 3: Blueberry polyphenols extend lifespan and slow aging in C. elegans
Literature Review
Figure 4: Structural and functional declines in C. elegans pharynx from days 2-10.
Chow et. al. 2006
Literature Review
Figure 5: Visible Structural Decline in C. elegans as a result of Sarcopenia Herndon et. al. (2002)
PurposeTherefore the purpose of this experiment is to test the ability of different concentrations of fruit polyphenols on the prolonging of lifespan in C. elegans.
HypothesisH(0): The pelargonidin chloride will show no effect on the prolonging of lifespan in C. elegans.H(0): The cyanidin chloride will show no effect on the prolonging of lifespan in C. elegans
H(a): The pelargonidin chloride will show a positive increase in the lifespan of C. elegans.H(a): The cyanidin chloride will show a positive increase in the lifespan of C. elegans.
Methodology
Caenorhabditis elegans
Control n = 150
Pelargonidin Chloride 200 ug/mln = 50
MeasurementAging of C. elegans measurement possible with the use of FUDR.
Statistical AnalysisANOVA run by SPSS followed by a Scheffe Post Hoc Test
Pelargonidin Chloride 100ug/ml n = 50
Pelargonidin Chloride 50ug/mln = 50
Cyanidin Chloride 200ug/mgn = 75
Cyanidin Chloride
50ug/ml
N= 75
Pelgargonidin ChorideN = 150
Cyanidin ChorideN = 150
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
BudgetVendorVendor Catalogue Catalogue
##PagePage ItemItem Item #Item # QuantiQuanti
tytyUnit $$Unit $$ Total $Total $
$$
Carolina Carolina Science 2009Science 2009
7878 248248 C. C. eleganselegans N2 (wild N2 (wild
type)type)
NP-17-NP-17-35003500
11 $10.95 (per $10.95 (per culture)culture)
$10.95$10.95
Caroline Caroline Science 2009Science 2009
7878 248248 NematodNematode Growth e Growth
AgarAgar
NP-17-NP-17-35203520
55 $6.25 (per $6.25 (per 135 ml 135 ml bottle)bottle)
$31.25$31.25
https://https://www.waters.cwww.waters.c
om/waters/om/waters/partDetail.htmpartDetail.htm
??partNumber=partNumber=WAT066224WAT066224
n/an/a n/an/a SymmetrSymmetry C18 y C18
Column, Column, 3.5 µm, 3.5 µm, 4.6 x 75 4.6 x 75
mmmm
WAT06WAT0662246224
11 $543.00$543.00 $543.0$543.000
Carolina Science Website
n/an/a n/an/a MethanolMethanol 874950 11 $5.75 per $5.75 per
500 mL500 mL$5.75$5.75
Total Price: $590.95
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