Jonathon Jackson and Dr. Robert Seegmiller, Zoology
As has been outlined, I am submitting this report of the accomplishments that have been made over the past semester in conjunction with the 1,000 dollar award that I received from your office.
First, I would like to thank those people who have made financial contributions and those who have given of their time to support the undergraduate research program. I recognize that the university makes an investment by offering these scholarships; it is a well-made investment.
I have spent the past semester working in a research laboratory with Dr. Robert Seegmiller of the department of Zoology and with James Pace, a graduate student. I have been involved with Dr. Seegmiller’s research in teratology, particularly the study of genetic mutations resulting in dwarfism. These mutations are possibly linked to arthritis. This letter reports on my experience. My report includes a review of the objectives that I stated in my proposal submitted last fall, a summary of what I was able to accomplish, and a statement on what I feel the benefits of the project have been for the laboratory and for myself.
I here briefly review the nature of the study I have been doing. I have helped in the curation of two different colonies of laboratory mice. Each colony has a mutant gene which shows a propensity for dwarfism. (The conditions are called chondrodysplasia, abbreviated cho, and Disproportionate Micromelia, abbreviated Dmm.) We have studied these mutations during the semester.
As I outlined in my proposal, I had the following objectives for my work with Dr. Seegmiller.
1. Complete genotyping of the cho colony, and explore alternate methods of this genotyping for future use.
2. Mate a heterozygote from each colony of mice to produce offspring that are heterozygous at both alleles.
During the four months of the semester, we have been able to accomplish several things. Some of the objectives mentioned above were completed, others were not.
The work I have done over the semester has included genotyping assays as well as animal curation. We have used two assays to map the gene sequence of the animals. The first is the Polymerase Chain Reaction, in which a strand of DNA is copied multiple times so that its base pair sequence can be determined. The second is electrophoresis, an assay which makes use of an electric current to differentiate between different molecules (in our case, between mutated and normal DNA). You will find enclosed a copy of the genotyping protocol which I have written up. My curation work has involved tagging and segregating mice. I have also taken tissue samples from which we have isolated DNA.
To meet the first objective, we spent a large amount of time working through the genotyping of the cho colony. This has been the major project I have been involved with. I have worked in conjunction with James Pace. The cho colony was overpopulated, and none of the animals had confirmed genetic makeup. We spent three months attempting to successfully genotype the colony. We had difficulties with the gel electrophoresis assay which shows whether or not a selected portion of DNA is mutated. The results of our experiments were not conclusive enough to confirm genotype. We suspect that the problem was due to the materials we used to make up the solutions for electrophoresis (the DNA primers to be specific). After multiple runs without success, we had the procedure performed by a colleague of Dr. Seegmiller. This person was able to conclusively identify the heterozygous animals in the colony.
Once we obtained this information, we were able to reduce the size of this colony and begin breeding between heterozygous animals. Although we were not able to genotype this colony, with help from this colleague we obtained the genotypes, and were able to isolate all the animals which carry the gene under study. The colony is now organized and in a position to be further studied. Thus, I have been able to help in the genotyping and cleaning up of this colony of laboratory animals, and further study is possible because of this work.
We have just begun the same process for the Dmm colony. Because of the amount of time involved in getting the other colony up and running, the second objective (that of studying offspring of a parent from each colony) was not accomplished.
In summary, then, only one of the two objectives was met. Because of the unforeseen challenges in accomplishing the first goal, there has not been time to accomplish the second. Nonetheless, the work that has been done is substantial and contributes to accomplishing the bigger goal. This is to continue the studies of these two genetic mutations linked to dwarfism, and hypothetically arthritis.
I feel that I was able to help the laboratory by reducing the cho colony to a workable size and by beginning breeding between animals in the colony of known genotype. The lab’s curation costs will be lower, and more importantly further studies can be done because the genotypes of the animals are known.
I have benefitted a great deal from the research experience this semester. I have learned new techniques and gained information that will be useful in other research pursuits. This is inclusive of both the techniques I have helped with, PCR and electrophoresis, as well as the aspects of animal curation I have learned. Dr. Seegmiller and James have helped me to better understand the role of research in science.
In conclusion, I have been able to accomplish one of two objectives chosen at the beginning of the project, and more importantly, I have benefitted from the educational experience. I feel the lab has also benefitted from the work I have done. This has been possible because of the funds received as a scholarship from your office, and I thus thank you for the reward.