Blog #2: Prohibitin and Mitochondrial Significance for Viable Progeny in Caenorhabditis elegans

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.

Blog #1: Prohibitin and Mitochondrial Significance for Viable Progeny in Caenorhabditis elegans

The title of my project is Prohibitin and Mitochondrial Significance for Viable Progeny in Caenorhabditis elegans. Dr. Marcello and I are studying the prohibitin protein complex (PHB) which is known to maintain the stability and function of mitochondria. Mitochondrial function is essential to having sufficient energy produced within cells as well as being linked to high fertility rates in humans. Accumulation of mutations in mitochondrial DNA is correlated with poor human egg quality as an adult female ages. C. elegans is an excellent model organism to study the function of this complex and its relationship to mitochondria as the nematode is transparent, small, and easily maintained. It is also excellent to study fertility and development as it has about a two week long lifespan and a single worm can produce between 300 to 1000 progeny. This research is important because the results will provide more information about the interactions and regulation of genes necessary for the maintenance of mitochondria. This in turn will can provide context for more effective treatment options created for mitochondrial dysfunction and infertility in humans.

My methods will include RNA interference where I will knocking out genes associated with the prohibitin protein complex, such as PHB-1 and PHB-2, then observe physical changes in the nematodes and progeny production. I can count the eggs and larvae produced on the agar plates and compare differences in these mutant nematodes and wild type nematodes. Using MitoTracker and splicing techniques, I can observe the physical changes in mitochondria morphology as a result of knocking out the different genes. Additionally, I can change the times of treatment length to observe for any significant variations in C elegans reproductive capabilities.

With these methods we hope to better understand the effect prohibitin and mitochondria have on developmental abilities in C elegans. I am thrilled to work on this project and present my findings at the end of the year. I hope to learn more about managing the research process and how to balance research with other academic and personal responsibilities throughout this academic year.