Conclusions and Next Steps
Conclusory Results
In conclusion, we were able to establish some significant differences in the behavior of VglutGAL4 flies as they age. As we saw last semester, the flies perform locomotor tasks worse as they age. We were unable to demonstrate that TP43 flies have reduced locomotor capabilities, which indicated that we need to perform more trials with these flies, or consider altering our methods of measuring locomotor deficiency. We were able to complete a trial with our Experimental TIMP flies, however they are not included here as we did not have a enough time to collect data with different groups of flies or different ages.
As so, without proper testing and further results we can't necessarily conclude anything at the moment. However we hypothesize that with more trials and testing, flies with an over-expression of TIMP1 can highlight a communication block between neurons and the extracellular matrix and overall worst locomotor behavior as seen in our results.
Next Steps
In preparation for next semester, we are hoping to perform more locomotor behavior tests to support our proposed conclusory result. Therefore, by acknowledging and applying accordingly the following limitations, we aspire to further our contributions to ALS research.
Some limitations for our study were:
Lack of flies for our tests; minimum was around 20 flies per test, however for some of our tests we were only able to test 7 to 15 flies.
Being unable to flip flies consistently. Due to our groups restricted schedules, we were unable to flip flies on a consistent schedule to ensure accurate age ranges in regards to locomotor testing.
Possible human-made errors as there could've been incorrect labeling on certain vials or wrongly identifying males and females in lab.
Acknowledging that drosophila isn't a perfect model for ALS' testing and that it won't perfectly translate to ALS' testing in humans.
At the moment we have pursued all three genes that were from our dataset to curate our results. However due to minimal testing and overall lack of time, we hope to continue testing our current genes (COMP, VAMP7/8, and TIMP) as our genes of interest to refine and/or further support our results.
Future Experimentation
For the upcoming semester we hope to set up these future crosses to further explore ALS' in drosophila and apply these findings to the scientific world.
Next semester, we are hoping to continue running locomotor trials on our flies that have overexpression of TIMP1. Currently we only have technical replicate trials of these flies, so next semester we would like to establish more biological replicate trial as well as investigate the impact of aging with this genotype. We would also like to set up a second group of TIMP flies that have expression on the X chromosome, because TIMP has been observed to have a differential expression depending on the location of the gene.
For our VAMP and TSP crosses, we will need to follow our cross set up utilizing double balancers. We have not gotten very far in this process due to the new gene stocks not producing progeny until the end of the semester. We hope to eventually have a stock of experimental flies for each of these genes, that have lowered expression of the gene in an ALS background.