So, six weeks in and I am finally getting a good roll on my project. I have my mutations which will introduce all cysteines into the beta subunit (D192CD193C) or all aspartate into the alpha subunit (C192DC193D) of the acetylcholine receptor. This project is actually a lot of fun for me to learn about because everything is a new skill to me. My mentor has set a task for me to collect dosage responses and desensitization data. I am very excited to start using the Picospritzer, which is a different method of acetylcholine (Ach) application.
I am working on a Two Electrode Voltage clamp (TEVc) rig. It is quite difficult to learn how to operate with two electrodes but it is most gratifying to successfully impale an egg and get current. This method washes the Ach over the egg through an injector line. The Picospritzer is another glass needle that is separate from the TEVc and is used to puff the Ach directly over the egg for more accurate current readings. I am still struggling with the learning curve but I am ready to impale and get my data.
The overall goal of my project is to characterize these mutations and their effect on the Ach response. Every day is a new set of challenges but the end is going to be worth the journey. I am looking forward to my last few weeks of INBRE and what my data reveals about the mutations in the Ach receptor.