By: Ashley Haugerud & Leah Cox
26 APRIL 2019
Background
Our research is based on the Anolis carolinensis, also known as the green anole lizard. This species can be found in the southeastern part of the United States such as Georgia, South Carolina, etc.The behaviors of the breeding season (May) and non-breeding season (October) of the lizards are very different, with the breeding season for the green anole usually starting in mid-April and lasting until August. The males will extend their brightly-colored dewlap to attract the female who they wish to mate with. For our research stream, we are looking at the reproductive behaviors of the lizards specifically in the breeding and non-breeding seasons, and what genes have an important role in controlling these behaviors. To begin our research, we chose the NPAS2 (neuronal PAS domain protein 2) gene in the Anolis carolinensis. This gene could potentially have a role in reproduction and behavioral differences of the green anole lizards.
Research
Previously, after we chose our gene, we picked our primers that we thought would be effective in our research. Some of the procedures that we conducted in the past include reconstitution of our primers, PCR, gel electrophoresis, gel imaging, and PCR cleanup. The most recent experiment that we have conducted is RNA isolation. The purpose of RNA isolation is to extract the mRNA to determine how the gene is expressed at a specific time in both the breeding and non-breeding season. We started RNA isolation by doing a few practice runs with liver tissue to be more prepared for the tissue we are studying, which is brain tissue. In previous experiments the first practice run we preformed went well, but we still needed some more practice. This is why we did another practice RNA isolation and the results were more efficient. We knew that we had good results because we obtained two clear bands on our gel image, and the purity (~1.8) of the RNA matched the standard purity. Two clear bands on a gel image means that our RNA is efficient and not degraded. After we performed two practice runs, we felt we had enough experience to move on to the brain tissue. We have performed five RNA isolations on the brain tissue. For two samples, we obtained good results and were satisfied with them. For the one sample, we did not get good results. Since the results were not satisfactory, we ran another gel. Before running the gel, we knew that we had two explanations as to why we did not get bands. The first reason is because the gel may not have worked. The next reason is because our RNA was possibly degraded. Currently, we are performing RNA isolation for sample 30 and 35. This involves the same process as the other RNA extractions. We obtained two clear bands in sample 30, but the two bands for sample 35 were faint. This could be due to a problem in the gel imaging process. We are also working on our final presentation for this research stream.
Next Steps
Next, we will continue with RNA isolation for our remaining samples and re-run the gel for sample 35, as well as work on our final presentations to show the results of our research from the past semester of experiments. Once we are finished with RNA isolation for all of our given brain tissue samples, we will continue research on how the NPAS2 gene is expressed in the brains of Anolis carolinensis in the breeding and non-breeding seasons.
References
“Green Anole (Anolis carolinensis).” Savannah River Ecology Laboratory University of Georgia. https://srelherp.uga.edu/lizards/anocar.htm
“Carolina Anole or Green Anole.” BioExpedition. May 9, 2012.https://www.bioexpedition.com/carolina-anole-or-green-anole/
