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

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Presentation on theme: "Resveratrol and Decreased Fat Deposition in C. elegans Alexandra Golway- Period 3 Manhasset Science Research."— Presentation transcript:

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

2 1998 Obesity Trends* Among U.S. Adults 1990, 1998, 2007 (*BMI 30, or about 30 lbs. overweight for 54 person) No Data <10% 10%–14% 15%–19% 20%–24% 25%–29% 30% Figure 1: Obesity trends among U.S. adults Source: Center for Disease Control & Prevention [http://www.cdc.gov/nccdphp/dnpa/obesity/trend/maps/obesity_trends_2007.ppt#487,5,Slide 5] Need

3 Knowledge Base Figure 2: 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]

4 Resveratrol Figure 3: chemical structure of resveratrol (C 14 H 12 O 3 ) [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

5 Literature Review RNAi Mutant C. elegans strains Nile Red Staining ingested Fat deposition monitored as RNAi silences genes Results 305 gene silences cause reduced fat storage 112 gene silences cause increased fat storage Wild-type C. elegans Figure 4: 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]

6 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 dauer organisms Wild-type N2 strain- Has AMPK Sudan Black staining Fat reserves Rationed at Constant level aak-2 mutant strain No AMPK Sudan Black staining Fat reserves destroyed Organisms died Figure 5: The enzyme AMPK is necessary for C. elegans in dauer to survive and regulate fat deposition (Roy, et al, 2009) [chart created by author]

7 Literature Review Resveratrol improves health and survival of mice on a high- calorie diet (Baur, et al, 2006) Positive effects of Resveratrol in mice Increased survival Activation of AMPK Increased insulin Sensitivity less fatty acid syntase More mitochondria Less damage to Liver and pancreas Figure 6: 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]

8 Literature Review Anti-obesity properties of resveratrol Stimulates adiponectin Decreased fat storage Human preadipocytes prevented from becoming fat cells Figure 7: 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]

9 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

10 Methodology C. Elegans N= 75 25= control- no resveratrol or caloric restriction 25= experimental- caloric restriction Sudan Black Staining Inserted into E. coli food- Ingested by organism Fluorescent microscope Examination- every 3 days Statistical analysis- Using SPSS program 25= experimental- Resveratrol ( 5uM,10uM, 20uM per day)

11 Protocols C. elegans placed in petri dishes Nematode Growth Medium (NGM) used E. coli is food source Resveratrol and Sudan Black will be ingested by organisms Picture by Victoria Wei

12 Budget VendorOrder #QuantityDescriptionUnit PriceTotal Price Caenorhabditis Genetic Center TJ Wild-type C. elegans N2$7.00$70.00 Sigma-AldrichR mg1Resveratrol- 100 mg$81.50 Carolina Biological 2009NP Sudan III Stain$26.85$53.70 Total Price: $205.20

13 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

14 Bibliography Ashrafi, K. Obesity and the regulation of fat metabolism (March 9, 2007),, ed. The Research Community, WormBook, doi/ /wormbook ,. 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): 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 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): The Endocrine Society. "Red Wine's Resveratrol May Help Battle Obesity." ScienceDaily 17 June June Roy, Richard and Patrick Narbonne. Caenorhabditis elegans dauers need LKB1/AMPK to ration lipid reserves and ensure long-term survival. Nature (2009) Vol Stiernagle, Theresa. Maintenance of C. elegans. February 11, 2006),, ed. The Research Community, WormBook, doi/ /wormbook ,.


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