Throughout the fall semester I have been working with Dr. Marcello to study prohibitin and mitochondrial significance in Caenorhabditis elegans. While waiting for the bacteria strain needed to perform the RNA interference (RNAi) experiments to arrive in the lab, I practiced my worm picking technique to preserve the wild type worms. Moving a few worms from one plate to a new one provided a diverse range of life stages for my lab mates to use for their experiments. My lab mates were an integral part of helping me perfect lab techniques and procedures. Additionally, it helped provide the necessary speed for my RNAi experiments because the special RNAi agar must be kept out of the light as much as possible. These experiments knock out the prohibitin gene so that I can observe the changes in progeny production in the mutant nematodes compared to the wild type.
When beginning the RNAi experiments, there were a couple problems we ran into. For example, condensation developing on some of the agar plates lead to contamination and chunking starved worms lead to a reduced number of viable worms in the L4 life stage. However, from the data we have collected, there does appear to be a difference in the amount of progeny produced by the mutants lacking prohibitin compared to the wild type worms. These experiments will need to be repeated in the future to ensure the validity of this finding.
After we have collected sufficient results from the RNAi experiments we hope to begin mitochondrial splicing in the spring semester. Prohibitin, the gene of interest, greatly affects mitochondria and we expect to see morphological differences in the mitochondria of the mutants lacking prohibitin compared to the wild type nematodes. Varying the treatment length of these experiments could possibly show significant variations in the reproductive capabilities of the nematode.