Methods
Virgin Collection:
Throughout the semester we collected virgins in order to make new crosses .
This is important because this is how we get new generations with wanted characteristics
To identify which were virgins we often looked at whether they were female, meaning if had stripes on their back (if they were males their back would be all black with no stripes) and if they had a black dot in their mid section.
If the females were no longer virgins then they would be used for the locomotor test
Locomotor Test:
This test was primarily us to test the mobility of the flies and to see how ALS affects their motor skills over time.
We used a graduated cylinder and out a yellow tape around the 100mL mark
We then transfered the flies from the vial into the graduated cylinder with the help of a funnel and covered it with parafilm so the flies would not fly away
We then tapped the graduated cylinder on the counter to make sure all flies start from the bottom.
Next we started a 2 minute timer and recorded as the flies flew up and passed the yellow tape.
After we carefully transfered the flies back into their vials for future testing.
Then took those videos and slowed them down to count exactly how many flies cross in 10 sec intervals and added that data to our spreadsheet.
Crosses:
Stocks:
we used these to get virgins in order to set up future crosses for behavioral testing
w1118; +/+; +/+
+/+; vglutGAL4/”; +/+
+/+; +/+; uas-TDP43/Tm6bSbTb
3-5 virgin females with 3-5 males
Crosses:
we began testing these genes as a set up to compare to our future gene D11
w1118; +/+; +/+ X +/+; vglutGAL4/”; +/+
+/+; vglutGAL4/”; +/+ X +/+; +/+; uas-TDP43/Tm6bSbTb
we take their progeny:
w1118; vglutGAL4; +/+
+/+; vglutGAL4; uas-TDP43/+
for this one specifically we looked out for ones that were not stubby or tubby because those did not have the uas- TDP43
for behavioral testing
Gene selection:
For our gene of intrest, D11, we selected this off of Dr. Alberto Catanese's data set after we were able to code his files and will set up crosses next semester.
Our Genes:
AP1M2: Enables Clathrin and Enables vesicle mediated transport between the synapses. Downregulated
MGST1: Helps to neutralize oxidative stress and prevents lipid peroxidation of membranes. Downregulated
FOXF1: Bind to serum response factor and myocardin regulating gene transcription in visceral smooth muscle. Upregulated
H2BC8: Involved in chromatin organization. Downregulated
Next page: Results and Conclusions