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. 2024 May 13;147(6):2053–2068. doi: 10.1093/brain/awae078

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© The Author(s) 2024. Published by Oxford University Press on behalf of the Guarantors of Brain.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Co-expression of RGNEF or NF242 with TDP-43 in fruit flies. (A) Kaplan-Meier graph showing the survival of elav>RGNEF (elav>R; n = 156), elav>NF242 (elav>NF; n = 101) RGNEF no driver control (R n.e.; n = 117), NF242 no driver control (NF n.e.; n = 93) and elav>w− (control of driver crossed with parental line; n = 123). The elav>RGNEF line shows an increased lifespan compared to RGNEF no driver control (P = 0.0035) and elav>w− (P < 0.0001) lines. The elav>NF242 line shows an increased lifespan compared to NF242 no driver control (P < 0.0001) and elav>w− (P < 0.0001) lines. (B) Kaplan-Meier graph showing the survival of elav>GFP;TDP-43^wt (elav>G; T; n = 178), elav>RGNEF;TDP-43^wt (elav>R; T, n = 132) and elav>NF242;TDP-43^wt (elav>NF; T, n = 224). The elav>GFP;TDP-43^wt line shows a reduced lifespan, an effect that is suppressed in the elav>RGNEF; TDP-43^wt (P < 0.0001) and elav>NF242;TDP-43^wt (P < 0.0001) lines. (C) Kaplan-Meier graph showing the survival of D42>GFP; TDP-43^wt (D42>G; T; n = 143), D42>NF242;TDP-43^wt (D42>NF; T, n = 181) and D42>w⁻ (control of driver crossed with parental line, n = 98). The D42>GFP;TDP-43^wt line shows a reduced lifespan, an effect that is suppressed in the D42>NF242 line (P < 0.0001). The latter also show an increase in lifespan compared to the control D42>w⁻ line (P < 0.0001). (D and E) Negative geotaxis assay showing the climbing score at Days 3 and 6 for elav>RGNEF;TDP-43^wt (elav>R; T, n = 11; 110 flies), elav>NF242;TDP-43^wt (elav>NF; T, n = 12, 120 flies), elav>GFP;TDP-43^wt (elav>G; T; with n = 12; 120 flies at Day 1) and elav>w⁻ (n = 8; 80 flies) lines. The elav>GFP;TDP-43^wt line shows a severe motor phenotype that is suppressed when RGNEF or NF242 is co-expressed with TDP-43^wt in neurons (P < 0.0001). (F and G) Negative geotaxis assay showing the climbing score at Days 6 and 9 for D42>RGNEF;TDP-43^wt (D42>R; T, n = 12; 120 flies) and D42>NF242;TDP-43^wt (D42>NF; T, n = 16, 160 flies), D42>GFP;TDP-43^wt (D42>G; T; n = 14; 140 flies) and D42>w⁻ (n = 9; 90 flies). The D42>GFP; TDP-43^wt line shows a significant motor phenotype that is suppressed when RGNEF or NF242 is co-expressed with TDP-43^wt in motor neurons (P < 0.0001). (H) Representative images showing the eye phenotype of GMR>w⁻ (negative control), GMR>36R (positive control), GMR>GFP;TDP-43^wt and GMR>NF242;TDP-43^wt lines. NF242 co-expression with TDP-43^wt suppresses the eye degeneration observed in the GMR>NF242;TDP-43^wt line. (I) Immunofluorescence of adult elav>NF242;TDP-43^wt fly brain tissue showing the co-localization between NF242 and TDP-43^wt in neurons. (J and K) Confocal images at higher magnification of adult elav>NF242;TDP-43^wt fly brain tissue showing the co-aggregation between NF242 and TDP-43^wt in neurons. Nuclei are indicated with dashed lines. Arrows show nuclear co-localization and arrowheads cytoplasmic co-localization.