1. Greater Understanding of Lipids in Caenorhabditis elegans Exposed to Oxygen Deprivation Padilla Lab, Department of Biological Sciences Jemma Alarcón TAMSter Researcher

2. Health Problems Related to Oxygen Deprivation Myocardial infarction Myocardial infarction Stroke Stroke Shock due to low blood pressure caused by vascular disease, inadequate heart function, or low blood volume Shock due to low blood pressure caused by vascular disease, inadequate heart function, or low blood volume Solid tumor cell resistance to radiation and chemotherapy Solid tumor cell resistance to radiation and chemotherapy Pulmonary disease and dysfunction Underdeveloped lung function in premature babies Anemia due to blood loss from severe injury or destruction of red blood cells Stroke alone accounts for 1 out of every 15 deaths in the United States and is the third leading cause of death in most developed countries.

3. Research Approaches to Studying O 2 Deprivation Physiology Physiology - Sea turtles - Brine Shrimp - Diving Seals - Birds Cell Biology Cell Biology - Cell culture Genetic approach Genetic approach - Drosophila melanogaster “Fruit flyes” - Caenorhabditis elegans 3 http://www.acuariolasmercedes.com/Guia-de-cuidado/cria-de-artemia-salina.htm 4 www.exploratorium.edu/.../ mutant_flies/ebony.gif 1 www.scubamaui.com/ turtles.htm

4. Anoxia-Induced Suspended Animation Normoxia Anoxia Time 0 24 hours Anoxia (<.001kPa oxygen in environment) Padilla, 2002 Mol. Biol. Cell, May 13(5) Embryos and larvae were exposed to either normoxic or anoxic conditions

5. Question and approach to Evaluating lipids in C. elegans exposed to anoxia. Hypothesis: Oxygen deprivation affects lipid accumulation. Hypothesis: Oxygen deprivation affects lipid accumulation. Approach: Use the dye Nile Red to assay lipid/fat accumulation in C. elegans. Approach: Use the dye Nile Red to assay lipid/fat accumulation in C. elegans. –In C. elegans Nile Red is taken up and attaches to lipids in intestinal cells. Lipids are visualized using fluorescent microscopy. Results: Increased detection of lipids in worms exposed to anoxia. Results: Increased detection of lipids in worms exposed to anoxia. –This result leads to the question of whether lipids are important for anoxia survival. Ashrafi, K et al, (2003). Nature 421, 268-272.

6. DIC Nomarski Microscopy

7. Fluorescent microscopy can be used to assay lipids via Nile Red detection Lipid Droplets

8. Normoxia 1 Day Anoxia 2 Days Anoxia DICNile Red Nematodes Exposed to 1 or 2 days of Anoxia Accumulate Lipids as Seen by Nile Red Detection. Size bar equals 100µm.

9. Comparison of Lipid Accumulation in Wild type (N2) and daf-2 Strains Normoxia 1 Day Anoxia 2 Days Anoxia N2daf-2 Size bar equals 100µm.

10. Discussion and Conclusion Lipid accumulation - Measure intensity - RNAi experiments - Viability Continue with daf-2 experiments - RNAi experiments - Viability

11. Acknowledgments Project was supported in part by a grant from National Institute of Health TAMS Summer Research Scholarship. Special thanks to Dr. Padilla for mentoring this project. I thank Dr. Padilla lab members for valuable input and suggestions. Alex Mendenhall Neil Stewart Vinita Hajeri Sara Anvari Bobby LaRue Nicole Parker Jesus Trejo! Desh Mohan Worms rule! http://www.celegans.de/ http://www.bio.unc.edu/ahmed/lab/ Dr. Padilla