Blog #3: Determining the function of F10C2.4 in reproduction

Since my last blog post, I have made promising progress with my research project. My main goal of this semester was to focus on microscopy. Now that I have established that my gene of interest, F10C2.4, clearly shows some kind of reproduction problem when not present using RNA interference, I wanted to utilize the unique feature of my model organism, Caenorhabditis elegans, to determine what is going wrong in the organisms without this gene. One of the drawbacks from using technology such as a microscope hooked up to a computer is that sometimes things can go wrong; however, I feel that I have learned so much from the mistakes, which has helped me get a better understanding of the research I do.
Caenorhabditis elegans have many benefits for genetic manipulation and research. One of the most beneficial features is that it is transparent. This is great for microscopy because it makes it easier for us to see what is different with the worm’s reproductive system when comparing it to the normal, not treated worm. For the experiments I perform for the microscopy element, we repeat the RNAi interference experiments with strains with fluorescent markers. GFPs are green fluorescent proteins that can stain a particular part of a cell; like a cell wall and RFP are red fluorescent protein can stain the chromosomes within the nucleus of the cell. With the strain I am working with, AJ740, I can utilize the GFP and RFP to see what is happening to the shape and overall placement of the eggs within the affect mother worm treated through RNA interference along with what is going on with the chromosomes. I have several questions. General questions like: are the eggs going to the right place and are there too many or too little eggs in the mom’s body? In regards to the chromosomes, are they separating at the right time? Is there an issue that is causing the incorrect amount of chromosomes in the cell which is not allowing them to hatch once they are laid? Does the egg go through the correct amount of cell divisions before being laid? All these questions can be answered using microscopy thanks to the organism, C. elegans.
A challenge that I have been facing, along with other members in my research group, is in regards to getting the microscopy aspect to work. This has opened my eyes to the many components involved in a task like this where things can go all good for one week but the next week, multiple things can go wrong. That being said, it is important to troubleshoot what went wrong and come up with ways to fix the problem at hand. An example of this is when the GFP and RFP do not show up bright enough to identify anything going on in the C. elegans. This could be due to the organisms not growing properly due to a lack of food or temperature sensitivity. Whichever the case, it was important for us to work together as a team to come up with a way to avoid this issue for the next week. For the future, I hope I can take more images from microscopy to look further into what is going on internally to determine what the function is of my gene of interest, F10C2.4.

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