1 / 19

The Effects of PUGNAc on O-GlcNAC Levels and Dauer Formation in C. elegans

The Effects of PUGNAc on O-GlcNAC Levels and Dauer Formation in C. elegans. By Nicole Kiess. Need. Picture 1 : Shows the rate of Type 2 Diabetes in relationship to obesity over 10 years. http://www.ndep.nih.gov/diabetes/WTMD/images/figure2.gif.

Ava
Download Presentation

The Effects of PUGNAc on O-GlcNAC Levels and Dauer Formation in C. elegans

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. The Effects of PUGNAc on O-GlcNAC Levels and Dauer Formation in C. elegans By Nicole Kiess

  2. Need Picture 1: Shows the rate of Type 2 Diabetes in relationship to obesity over 10 years. http://www.ndep.nih.gov/diabetes/WTMD/images/figure2.gif

  3. Estimated prevalence of diagnosed and undiagnosed diabetes in people aged 20 years or older, by age group, United States, 2007 Need Picture 2: Shows percent of people in different age groups with Type 2 Diabetes. http://www.cdc.gov/diabetes/pubs/estimates07.htm

  4. Knowledge Base Picture 3: Is a model of what happends in the body of a Type 2 Diabetic http://savvyhealthfitness.com/get-healthy/diabetes/type-2-diabetes-lifestyle-diabetes

  5. Knowledge Base Picture 4a: Shows the pathway that regulate aging in C. Elegans. Picture4 b: Shows the model for the regulation of life span and sensory perception. http://www.nature.com/nature/journal/v408/n6809/fig_tab/408255a0_F3.html

  6. Knowledge Base Picture 5:C. Elegnas life cycle. http://www2.niddk.nih.gov/NR/rdonlyres/6B1E037B-3958-485C-8CFE-A448E43814DD/0/dauer2.jpg

  7. Literature Review • Paradis, et al,(1998) showed that mg144 does not activate AGE-1 P13K and that Akt/PBK is not dependent on AGE-1/P13K Picture 6a: Shows normal growth conditions Picture 6b: Shows dauer formation conditions

  8. Literature Review • Arias, et al, 2004 showed that PUGNAc increases O-GlcNAc levels in rat skeletal muscle Picture 7: Rate of 3-MG transport with or without PUGNAc. Rats were given different concentrations of insulin .

  9. Literature Review • Hanover, et al, 2005, findings show that OGT modulates macronutrient storage and dauer formation in C. Elegans Picture 8: Shows percentages of dauers in different strains

  10. Literature Review • Forsythe, et al, 2006 demonstrated that enzymes of O-GlcNAc cycling perfect insulin-like signaling in response to nutrient flux Picture 9: Dauer formation in different strains with temperature as a variable.

  11. Purpose • Therefore the purpose of this experiment is to observe the effects of PUGNAc on O-GlcNAc levels and dauer formation levels

  12. Hypothesis • Null- There will be no change in O-GlcNAc levels and no change in dauer formation • Alternative- The levels of O-GlcNAc will rise and dauer formation will rise

  13. Method The effects of PUGNAc on O-GlcNAC levels and dauer fomation in the insulin-like signaling pathway of C. elegans Control Wild Type N2 Ogt-1 (ok430) Oga-1 (ok1207) Daf-2 (e1370) 50um 100um 50um 100um 50um 100um 50um 100um • C. Elegans will be cultured at 16 degrees C • O-GlcNAC levels will be observed by treating C. elegans with RL1 and RL2 antibodies • Pictures will be taken with camera under immunofluorescent microscope and photoshop will measure percentages • Adults of each strain lay embryos overnight at 15 degrees C. Cultures shifted to 22.5 degrees C and are scored 3 days later as either dauer or non dauer Statistical Analysis will be run with SPSS and an ANOVA

  14. Protocol • C. eleganswill be cultured in petri dishes with the Nematode Growth Medium (NGM) and will be fed Escherichia coli (E. coli) Picture 10: How the C. elegans will be cultured Drawn by Victoria Wei

  15. Protocol • RL1 and RL2 dilated 1:400 and given to C. Elegans Picture 11: Life cycle of C. elegans http://www.sfu.ca/biology/faculty/hutter/hutterlab/research/pictures/Ce_life_cycle.gif

  16. Protocol • Fluorescent microscope and camera used for picture • Photoshop used for percentages Picture 12: A) Fluorescent picture to show O-GlcNAc levels with RL-2 B) Fluorescent picture to show O-GlcNAc levels with RL-2 Hanover, et al, 2005

  17. Do-Ability • Need to Purchase: • RL-1 RL-2 • C. elegans strains • E. coli • PUGNAc • Already Obtained • Fluorescent microscope • Camera • Photoshop • Petri Dishes

  18. Budget

  19. Bibliography • Biggs, Alton, Whitney Crispen Hagins, Chris Kapicka, Linda Lundgren, Peter Rillero, and Kathleen G. Tallman. Biology The Dinamics of Life. Columbus, Ohio: The McGraw-Hill Companies Inc., 2004 • Cauter, Eve V., Rachel Leproult, Esra Tasali, and Karine Spiegel. "Effects of poor and short sleep on glucose metabolism and obesity risk." Nature Reviews Endocrinology 5 (2009): 253-61. • Chang, Annette M., and Jeffrey B. Halter. "Aging and Insulin secretion." AJP-Endocrinol Metab 284 (2003). • Clevenger, Christopher M., Pamela Parker Jones, Hirofumi Tanaka, Douglas R. Seals, and Christopher A. DeSouza. "Decline in insulin action with age in endurance-trained humans." Journal of APplied Physiology 93 (2002): 2105-111. • "Introduction to C. Elegans." Genes, Genomes, and Human. 07 May 2009 <http://avery.rutgers.edu/WSSP/StudentScholars/project/introduction/worms.html>. • Kao, G., C. Nordenson, M. Still, A. Ronnlund, S. Tuck, and P. Naredi. "ASNA-1 positively regulates insulin secretion in C. elegans and mammalian cells." Cell 128 (2007): 577-87. • Kimura K.D., Tissenbaum H.A., Liu Y., Ruvkun G. (1997) daf-2, an insulin receptor-like gene that regulates longevity and diapause in Caenorhabditis elegans.Science277:942–946. • Kodama, Eiji, Atsushi Kuhara, Akiko Mohri-Shiomi, Koutarou D. Kimura, Masatoshi Okumura, Masahiro Tomioka, Yuichi Iino, and Ikue Mori. "Insulin-like signaling and the neural circuit for integrative behavior in C. Elgans." Genes and Development 20 (2006): 2955-960. • Manzella, Debra. "Obesity and Type 2 Diabetes - Why Being Overweight Can Damage Health." Diabetes - Diabetes Management - Signs of Diabetes - Symptoms of Diabetes. 04 May 2009 <http://diabetes.about.com/od/symptomsdiagnosis/a/obesity.htm>. • Muzumadar, Radhika, Xiaohui Ma, Gil Atzmon, Patricia Vuguin, Xiaoman Yang, and Nir Barzilai. "Decrease in Glucose-Stimulated Insulin Secretion With Agine Is Independent of Insulin Action." Diabetes 53 (2004): 441-46. • Norman, James. "Introduction to Insulin: insulin discovery and actions." Endocrine Diseases: thyroid, parathyroid adrenal and diabetes. 07 May 2009 <http://www.endocrineweb.com/diabetes/2insulin.html>. • Paradis, Suzanne, and Gary Ruvkun. "Caenorhabditis elegans Akt/PKB transduces insulin receptor-like signals from AGE-1 P13 kinase to the DAF-16 transcription factor." Genes and Development 12 (1998): 2488-498. • "Type 2 Diabetes: Conditions, Treatments, Resources - American Diabetes Association." American Diabetes Association Home Page. 04 May 2009 <http://www.diabetes.org/type-2-diabetes.jsp>. • Twyman, Richard. "Model organisms: The nematode worm." The Human Genome. 07 May 2009 <http://genome.wellcome.ac.uk/doc_WTD020809.html>. • Waksman Student Scholars. "Introduction to C. Elegans." Genes, Genomes, and Human Genetics. 12 May 2009 <http://avery.rutgers.edu/WSSP/StudentScholars/project/introduction/worms.html>.

More Related