3rd instar Drosophila larvae genotypes
OK371>Gal4/ UAS-Venus-TDP-43 WT, OK371>Gal4/ UAS-Venus-TDP-43
M337V, and OK371>Gal4/UAS-YFP-TDP-43 A315T were used for axonal transport
imaging. For TDP-43 quantification at the cell body and synaptic terminal,
UAS-CD8-RFP (Bloomington Stock Center, Indiana U, Bloomington, IN; stock #27391)
was co-expressed, resulting in the genotypes: OK371>Gal4, UAS-CD8-RFP/
UAS-TDP-43 WT or OK371>Gal4, UAS-CD8-RFP/ UAS-TDP-43 M337V,
OK371>Gal4, UAS-CD8-RFP/UAS-TDP-43 A315T, and negative control-
OK371>Gal4, UAS-CD8-RFP/+. OK371-Gal4 driver was used for all
Drosophila transgene expression except for the TBPH rescue
cross where Armadillo Gal4 was used. In TBPH viability rescue cross, transgenes
(UAS-Venus-TBPH, UAS-Venus-TDP-43, and UAS-Venus-TDP43 M337V) were expressed by
Armadillo Gal4 in TBPH Δ23 homozygote mutants. Controls in this analysis
included W1118, homozygotes and heterozygotes for the null TBPH Δ23
mutation. Third instar larvae were sorted by genotype and placed in separate
vials. The percentage of larvae that became living adults was recorded and
graphed.
Alami N.H., Smith R.B., Carrasco M.A., Williams L.A., Winborn C.S., Han S.S., Kiskinis E., Winborn B., Freibaum B.D., Kanagaraj A., Clare A.J., Badders N.M., Bilican B., Chaum E., Chandran S., Shaw C.E., Eggan K.C., Maniatis T, & Taylor J.P. (2014). Axonal transport of TDP-43 mRNA granules in neurons is impaired by ALS-causing mutations. Neuron, 81(3), 536-543.
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