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Resveratrol and Decreased Fat Deposition in C. elegans Alexandra ******- Period 3 Manhasset Science Research

Resveratrol and Decreased Fat Deposition in C. elegans

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Resveratrol and Decreased Fat Deposition in C. elegans. Alexandra ******- Period 3 Manhasset Science Research. Need. Obesity Trends* Among U.S. Adults 1990, 1998, 2007. (*BMI  30, or about 30 lbs. overweight for 5’4” person). 1998. 1990. 2007. - PowerPoint PPT Presentation

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Page 1: Resveratrol and Decreased Fat Deposition in  C. elegans

Resveratrol and Decreased Fat Deposition in C. elegans

Alexandra ******- Period 3

Manhasset Science Research

Page 2: Resveratrol and Decreased Fat Deposition in  C. elegans

1998

Obesity Trends* Among U.S. Adults1990, 1998, 2007

(*BMI 30, or about 30 lbs. overweight for 5’4” person)

2007

1990

No Data <10% 10%–14% 15%–19% 20%–24% 25%–29% ≥30%

Source: Center for Disease Control & Prevention [http://www.cdc.gov/nccdphp/dnpa/obesity/trend/maps/obesity_trends_2007.ppt#487,5,Slide 5]

Need

Page 3: Resveratrol and Decreased Fat Deposition in  C. elegans

Knowledge Base

Figure 1: Chemical structure of Nile Red (A) & Sudan Black (B) staining. Nile Red staining in wild-type and mutant C. elegans (C-E). Sudan Black staining in wild-type (F) & mutant (G) organisms.Source: (Ashrafi, 2007) [http://www.wormbook.org/chapters/www_obesity/obesity.html]

Page 4: Resveratrol and Decreased Fat Deposition in  C. elegans

Resveratrol

Figure 2: chemical structure of resveratrol (C14H12O3)

[http://www.3dchem.com/molecules.asp?ID=277]

• Chemical found in grapes; red wine

• Popular dietary supplement- advertised as anti-aging substance

•May explain “French paradox”

Page 5: Resveratrol and Decreased Fat Deposition in  C. elegans

Literature ReviewRNAi

Mutant C. elegans strains

Nile Red Staining ingested

Wild-type C. elegans

Fat deposition monitored as RNAi silences genes

Results

305 gene silences cause reduced fat storage

112 gene silences cause increased fat storage

Figure 2: RNAi was used to silence certain genes to determine which ones regulated fat in C. elegans (Ashrafi, et al, 2003) [chart made by author using materials from article]

Page 6: Resveratrol and Decreased Fat Deposition in  C. elegans

Literature Review

• C. elegans in dauer need AMP-activated protein kinase (AMPK) to survive– Dauer- larval “hibernation”

stage in which metabolic rate is slowed; fat deposits increase to sustain organism

– AMPK- fat regulatory enzyme

• aak2 mutant dauers lack AMPK destroy fat

• (Roy, et al, 2009)

C. Elegans dauerorganisms

Wild-type N2 strain-Has AMPK

aak-2 mutant strainNo AMPK

Sudan Blackstaining

Sudan Blackstaining

Fat reservesRationed at

Constant level

Fat reservesdestroyed

Organisms died

Figure 3: The enzyme AMPK is necessary for C. elegans in dauer to survive and regulate fat deposition (Roy, et al, 2009) [chart created by author]

Page 7: Resveratrol and Decreased Fat Deposition in  C. elegans

Literature Review• Resveratrol

improves health and survival of mice on a high-calorie diet

• (Baur, et al, 2006)

Positive effects ofResveratrol

in mice

Increased survival

Activation of AMPK Increased insulin

Sensitivityless fatty acid

syntase

More mitochondriaLess damage to

Liver and pancreas

Figure 4: Resveratrol caused many positive changes in the overall health of mice similar to the effects of caloric restriction

(Baur, et al, 2006) [chart made by author adapted from article]

Page 8: Resveratrol and Decreased Fat Deposition in  C. elegans

Literature Review

Human preadipocytes

prevented from becomingfat cells

Decreasedfat storage

Stimulates adiponectin

Anti-obesity properties

of resveratrol

Figure 5: In human preadipocytes (fat cell precursors), resveratrol in extremely high doses (3-5 g) caused positive changes in the cells. (Fischer-Posovsky, et al, 2008) [chart made by author]

Page 9: Resveratrol and Decreased Fat Deposition in  C. elegans

Purpose/Hypothesis

• Purpose: to observe the effects of resveratrol on fat deposition in C. elegans and compare it to caloric restriction

• Null hypothesis: Resveratrol will cause no change in fat deposition in C. elegans

• Alternate hypothesis: Resveratrol will cause a decrease in fat deposition and mimic the effects of caloric restriction

Page 10: Resveratrol and Decreased Fat Deposition in  C. elegans

Methodology

C. ElegansN= 75

25= control- no resveratrol

or caloric restriction

25= experimental- caloric restriction

25= experimental- Resveratrol

(10uM per day)

Sudan Black StainingInserted into E. coli food-

Ingested by organism

Fluorescent microscopeExamination- every 3 days

Statistical analysis-Using SPSS program

Page 11: Resveratrol and Decreased Fat Deposition in  C. elegans

Budget

Vendor Order # Quantity Description Unit Price Total Price

Caenorhabditis Genetic Center TJ3014 10 Wild-type C. elegans N2 $7.00 $70.00

Sigma-Aldrich R5010-100mg 1 Resveratrol- 100 mg $81.50 $81.50

Carolina Biological 2009 NP-17-3500 2 Sudan III Stain $26.85 $53.70

Total Price: $205.20

Page 12: Resveratrol and Decreased Fat Deposition in  C. elegans

Do-ability

• Resveratrol studies have been done in previous years

• C. elegans is a useful model organism that is easily obtained

• Sudan Black staining and fluorescent microscope examination are possible

Page 13: Resveratrol and Decreased Fat Deposition in  C. elegans

Bibliography• Ashrafi, K. Obesity and the regulation of fat metabolism (March 9, 2007), , ed. The Research Community, WormBook,

doi/10.1895/wormbook.1.130.1, <http://www.wormbook.org/chapters/www_obesity/obesity.html>.• Ashrafi, Kaveh, Francesca Y. Chang, Jennifer L Watts, Andrew G. Fraser, Ravi S. Kamath, Julie Ahringer, and Gary Ruvkin. “Genome-

wide RNAi analysis of Caenorhabditis elegans fat regulatory genes.” Nature (2003): 268-72• Baur, Joseph A., Kevin J. Pearson, Nathan L. Price, Hamish A. Jamieson, Charles Lerin, Avash Kalra, Vinayakumar V. Prabhu, Joanne Sl

Allard, Guillermo Lopez-Lluch, Kaitlyn Lewis, Paul J. Pistell, Suresh Poosala, Kevin G. Becker, Olivier Boss, Dana Gwinn, Mingyi Want, Sharan Ramaswamy, Kenneth W. Fishbein, Richard G. Spencer, Edward G. Lakatta, David Le Couteur, Reuben J. Shaw, Placido Navas, Pere Puigserver, Donald K. Ingram, Rafael de Cabo & David A. Sinclair. “Resveratrol improves health and survival of mice on a high-calorie diet.” Nature (2009) Vol. 444.

• Dasgupta, Biplab and Jeffrey Milbrandt. “Resveratrol stimulates AMP kinase activity in neurons.” Proceedings of the National Academy of Sciences of the United States of America. 2007, April 24: 104(17): 7217-7222

• The Endocrine Society. "Red Wine's Resveratrol May Help Battle Obesity." ScienceDaily 17 June 2008. 4 June 2009 <http://www.sciencedaily.com /releases/2008/06/080616115850.htm>.

• Roy, Richard and Patrick Narbonne. “Caenorhabditis elegans dauers need LKB1/AMPK to ration lipid reserves and ensure long-term survival.” Nature (2009) Vol. 457.

• Stiernagle, Theresa. “Maintenance of C. elegans.” February 11, 2006), , ed. The Research Community, WormBook, doi/10.1895/wormbook.1.101.1, <http://www.wormbook.org/chapters/www_obesity/obesity.html>.