Miura Wiley

Investigating the Role of cAMP in the Sleep Phenotype of the Drosophila Fragile X Syndrome Model

April 10, 2021   /  

Name: Miura Wiley
Major: Biochemistry and Molecular Biology
Minor: Spanish
Advisors: Dr. Seth Kelly, Dr. Rebecca Williams, Dr. James West (second reader)

Fragile X Syndrome (FXS) is the leading heritable cause of intellectual disability and is associated with a variety of neurological and behavioral phenotypes.  It has been linked with mutations in the FMR1 gene, which has an ortholog, dFmr1, in the model organism Drosophila melanogaster.  This model system has been used to identify the molecular basis through which the FMR1 gene exerts control over its associated phenotypes, such as memory, learning, and sleep processes.  One of the proposed molecular mechanisms is the cAMP signaling cascade, as null dFmr1 mutants are hypothesized to have decreased cAMP levels.  In order to investigate this mechanism, flies with mutations in the dFmr1 gene were crossed with dnc1 mutants, which are known to have increased cAMP levels.  This cross had previously been found to rescue a deficient sleep phenotype of dFmr1 mutants, and the proposed mechanism behind this restoration was through rescue of the cAMP cascade.  Here, the cAMP and sleep levels of both individual mutants as well as the crossed double mutant were measured to determine if cAMP levels were restored to wildtype levels in the cross.  No significant relationship was found between genotype and cAMP levels; however, this may be attributed to problems with the dnc1 genotype used in the cross.


 
Miura will be online to field comments on April 16:
10am-noon EDT (Asia: late evening, PST: 6-8am, Africa/Europe: early evening)

67 thoughts on “Investigating the Role of cAMP in the Sleep Phenotype of the Drosophila Fragile X Syndrome Model”

  1. Congratulations, Miura! I’m certain that you will be successful in your future endeavors.

  2. How can you have all this knowledge in your head, be a 2 sport stud-athlete, a great person and good friend ? You are an amazing example of making the most of your overall Wooster experience.

    1. Thanks Geordie, being able to play two sports and having such an amazing group of people to support me is the perfect example of #WhyDIII.

          1. What was your reasoning behind your hypothesis? Why did you think the cross between the dnc and fxs mutants would recur wild type cAMP levels?

          2. That’s a great question! In another study this cross restored wildtype sleep levels in the double mutant, and the predicted reason for this result was that cAMP levels were also restored to wildtype.

  3. Great work! You overcame some challenges with this project, but as Geordie stated, you made the most of the experience.

  4. Was there any observable consistency in your results for each of the genotypes between the ELISA and BCA assays?

    1. Hi Tom! There were consistent results for each of the genotypes in the sleep study, but there was a lot of variation found for the results of the ELISA and BCA assays.

  5. Super proud of your work Goose, so proud to have a scientist in our family and I know it’s been a rough COVID impacted year. We’re eager to see the presentation and understand the science behind it.

    1. Thanks Dad, I’ll put up a video of me giving the presentation and send you the link for that so you can watch the whole thing.

      1. Your subject is out of my knowledge base so I can’t directly address it but I want to say I know how difficult IS is, and I’m really impressed that you accomplished this while being such a driven and successful athlete, a winner of a person who gives so much to others, a true great representative of what The College of Wooster is all about! Congratulations!

        1. Thank you so much Ms. Thompson! I appreciate your support since you know about the I.S experience firsthand! I must say that your daughter being there for me on and off the field has made an immeasurable impact on my Wooster experience, so thank you for sending her my way!

    1. Good question, Jay! If I were to repeat the experiment, I would try to confirm the genotypes of all the flies prior to completing the molecular assays. I would also try to get a homozygous dFmr mutant to see how that compared.

  6. When discussing your future directions, how would you plan to confirm the genotype of the flies and what would it mean if the genotypes were correct but the assays produced the same results as your study?

    1. Thanks for the question. I would confirm the genotypes by completing the sleep study first to make sure they all match their expected sleep phenotypes. I would also probably look at PDE enzyme activity in the dnc1 mutants to make sure it is inhibited. If the genotypes were correct but the results were replicated, I would probably interpret that as a certain amount of variation being inherent to the genotypes.

  7. Congrats! This is amazing, when you present this information you are a natural. The world of genetics won’t know what hits them.

    1. Thanks so much E. Who would’ve thought a painter and a geneticist would make such a dynamic duo.

  8. Did this project make you more or less interested in the specific work done here? Do you see yourself pushing this particular research further or are you more likely to use the techniques and knowledge learned at Woo in other ways?

    1. This project definitely helped confirm my interest in the genetics lab work field in general, but I likely won’t continue with this specific topic. I really enjoyed the problem-solving aspect of this kind of work, and I think that that is something I will look for in whatever research I do moving forward. The rest of the techniques and knowledge I’ve learned at Woo will also be helpful in the genetics field regardless of the specific area of research, so it’s nice to have that kind of flexibility and freedom.

  9. This was an amazing study! I appreciated the connection made between the MGLUR and CAMP theories, which helped visualize your thinking process behind the experiment.
    One question I have about this experiment would be if you have considered testing mutated and non-mutated flies for memorization / pattern recognition differences instead of sleep?

    1. Thank you so much! Memorization, pattern recognition, and other measures of learning would be interesting to look at in these crosses, since we know that the two individual mutants have some deficiencies in these areas. That would be something great to test in future work, to see if the cross done here could rescue wildtype levels in these phenotypes as well!

  10. Proud of my dual sport scientist! I know how much hard work went into this project, especially overcoming the many setbacks you encountered. Looking forward to all the amazing things you will accomplish in your future!

    1. Thanks Mom, and thanks for always listening to me vent when said setbacks were encountered.

  11. Great job Miura! Those sleep results are interesting – definitely something to think about for the future!

    Thanks for all your hard work this year in lab and congrats on finishing I.S.!

    1. Thanks Dr. Kelly! Yes, the sleep results are definitely interesting. I’m glad you recommended this as the follow-up study since it really helped me interpret the molecular results. Thanks again for your guidance on this project!

  12. Nice, Muira! Your findings provide a great basis for further research in a field that is much needed. Congrats!

  13. Way to go miura!! Very proud (like everyone has said) that you made it through everything even though it didn’t work out the way you wanted it to. For those of us who know nothing about bcmb, what’s a sleep phenotype and what role did it have in your research?

    1. Thanks Lil, I can’t wait to see your awesome physics I.S. next year! A sleep phenotype is just a fancy way of saying an observable/measurable characteristic in the sleep physiology of these flies, so in my case, how much they slept overall, during the day, and at night. In my research, I used them to confirm the genotypes of my flies by comparing the expected sleep patterns to what I actually observed.

  14. What made you drawn towards Fragile X Syndrome and its proposed molecular mechanism of cAMP signaling?

    1. I was drawn toward FXS because it’s a common disease that affects so many people. I wanted to look at its molecular mechanism because if this aspect of the disease is understood, people can target it with therapeutic treatments in the future!

  15. Great presentation! As someone who is interested in genetics, I was wondering how you chose this topic and developed your research strategy for I.S.?

    1. Thanks Katie! I picked this topic since I was interested in the genetic basis of Intellectual Disability, and it’s an area of research that has so many aspects that still need to be explored. I developed my research strategy by doing a lot of reading and research on the background of Fragile X Syndrome since I initially didn’t know much about it. This helped me narrow my focus to one specific pathway associated with the disease that I could look at. I highly recommend doing work in genetics, it is an area where your research has the potential to ultimately help people!

  16. Also, CONGRATS Miura!! It’s so cool seeing my teammate doing big things and showing off how smart she is! It has been so fun to play with you on the field and now seeing what you have been doing in the classroom is so cool to hear about!

    1. Thanks Ann! I’m so glad I’ve gotten to know you on and off the field this season.

  17. Congratulations on all your hard work Miura! So proud to call you one of my closest friends. All your models look amazing. They truly convey the material clearly to the viewer.

    1. Thank you, Britta! I’m so lucky to have you in my life and I know you’re going to continue killing it at Woo the next two years.

  18. Congrats Miura! What do you think was the biggest challenge you had to overcome during this process?

    1. Thank you, Noah! The biggest challenge was probably trying to figure out the next step after getting the results for the molecular study. I had no idea how to interpret these findings, so I had to do some more research and prepare a follow-up study.

  19. Congrats Miura! Your work is incredible and I’m so proud of you for finishing. What do you think you will use the most out of this project in your future research?

    1. Thanks Maya! In the future I will definitely be using some of the problem-solving and troubleshooting skills I learned from this project, so it was a really valuable experience despite the challenges.

  20. I have such a smart teammate, roomie, and best friend!! So proud of you and everything you have accomplished and overcome throughout this project! So many amazing this to come 🙂

    1. Thank you so much, Ashley. I honestly couldn’t have done any of it without you, and I’m so lucky to call you my best friend <3

  21. Miura!

    Well done! I hope you are as happy with your work as I am and more.

    What a fun and productive time we had working together. I hope that our paths will cross again next when I’m on campus.

    Best wishes for a good celebration post IS,
    Dr. Williams

    1. Thank you, Dr. Williams! I had a great time working with you throughout this project, and I’m excited to work with you again next fall! Thanks again for all of your support.

  22. Congratulations Miura! You are finIShed. Proud of the hard work you’ve put into your project.

  23. Hi Miura!

    Great project, I liked the idea of using cross of two extreme phenotypes to potentially create a rescue phenotype. I have a question about the variability. As many can attest, IS is a learning experience and often are completing experimental lab techniques for the first time. Do you think the variability observed could be due to you “mastering” your technique. Also, did you think of any other ways to potentially study cAMP levels?

    Congrats on finIShing! I wish you all the best if you return to Wooster for a final semester and off in the future. If I can ever help you, let me know!

  24. Congratulations, Miura, on a very interesting and well-done study! Best wishes!

  25. Well done, Miura! It’s impressive to see how far you’ve come since FYS. Thanks for sharing this really interesting work.

  26. You’ve done a great job here Miura! Thank you for your encouragement in the lab this year, it’s wonderful to know someone like you took the time to chat with me. You’re going to wonderful things after Wooster!

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