Update 3: Wound Healing in Neural Cells

I am currently in my last week of research for the summer, trying to finish my examination of the three genes I’ve been working with. As I write this blog, I am running a third in situ hybridization with all of these genes: hdac7, psme3, and c9. The hdac7 probe I used worked in the last in situ, so I am hoping to confirm that the signal I saw last time will be repeated so I can be more confident about the expression pattern of this gene. I had all sorts of problems getting probes for the psme3 gene, which did not work in my last in situ — it took 4 attempts at a probe synthesis before I got results that look promising. However, there is no way to know for sure that the probes work without doing another in situ, so I will know in a day or so whether I actually succeeded. I synthesized the probes for the c9 gene without too many problems, but it is the first time I have tried an in situ with them as well. I am eagerly waiting to see the results of the color reaction that will show me where these three unique genes are expressed in an actual embryo, provided that everything works correctly. I am also learning how to cryosection embryos, a technique that allows me to get very detailed images of embryos that have undergone in situ hybridizations. Being able to prepare slides and get great photos of these embryos will help me to see more clearly exactly where my genes of interest show up in frog embryos, which is a big step towards determining what role they may play in wound healing. It isn’t the most enjoyable process, since it involves having my hands in -20 degrees Celsius for extended periods of time, but it will be worth it to learn more about these genes and get images that will look great in posters and papers in the future! I am definitely looking forward to continuing this research when I come back for the fall semester.

Comments

  1. gborgarelli says:

    Hi Regan, the work that you’re doing is fascinating! First of all, you’re a college student cloning genes, which sounds absolutely incredible. After reading all your blog posts it also sounds like a tedious and detailed job, but you seem to be handling it in the best way, learning from every success and mistake…so props to you! It sounds like your findings could have a major impact on wound healing in the future, and if I may ask, how did you think of connecting gene cloning and wound healing?

  2. rssindelar says:

    Hi! Thank you for the interest in my research project! The research lab that I work in on campus (Dr. Saha’s developmental biology lab) has multiple projects going on, and a former graduate’s honors thesis had to do with how embryos re-pattern and develop after ablation and rotation (meaning a piece of tissue was cut out, rotated, and placed back in the embryo). From his data, RNA sequencing gave a list of genes that were differently expressed in embryos that had been wounded and subsequently healed. Some of these genes were not well studied, so Dr. Saha, myself, and some other lab members became interested in studying their expression patterns in a normal embryo in order to see how they could be working in a healing embryo. In order to see expression patterns, the gene must be cloned in order to make many copies of RNA probes that bind to DNA in the embryo and signal exactly where the gene is “turned on.” Therefore, by cloning the genes, I was able to use in situ hybridizations with my probes, along with an antibody color reaction, to study expression patterns that could give an idea as to where and how these genes work in the healing process. I still have a lot to learn from this research, and I’m excited to build off of my summer project and keep exploring the complex process of wound healing and its many potential applications!

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