length human tau  (Addgene inc)

Journal: bioRxiv

Article Title: Injury-induced tau pathology promotes aggressive behavior in Drosophila without neurodegeneration

doi: 10.1101/2025.11.22.689595

Figure Lengend Snippet: Pan-neuronal human tau Experimental setup for collection and administration of injury to Drosophila virgin males. Days indicate post-eclosion. All experiments were conducted at 24 hours post-injury. B. Representative max projection images of whole brain immunohistochemistry using an antibody against human tau in UAS-Tau (control) and Elav-GAL4>UAS-Tau male flies for sham and injury conditions. Scale bars: 50 μm. C. Quantification of the percent of male flies with the indicated genotype and injury condition that climb 16 cm within 10, 15, and 20 sec in a negative geotaxis assay (n=8 trials ) . For UAS-Tau sham vs. UAS-Tau injury at 10, 15, and 20 seconds post-startle: p =0.0010, 0.0252, and 0.2415, respectively; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham at 10, 15, and 20 seconds post-startle, p <0.0001, p =0.0756, and p =0.1179, respectively; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury at 10, 15, and 20 seconds post-startle, p< 0.0001, 0.0219, and 0.0742, respectively; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury at 10, 15, and 20 seconds post-startle, p= 0.0769, 0.2951, and 0.5091, respectively. D-F. Noncompetitive courtship assays for UAS-Tau sham and injury male flies and Elav-GAL4>UAS-Tau sham and injury male flies (n=11, 9, 20, and 12 mating groups, respectively, from 3 independent trials). D. Quantification of the percent of fly pairs that mated within 2 hours. For UAS-Tau sham vs. UAS-Tau injury p= 0.4212; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.0284; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.6060; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0724; UAS-Tau injury vs. Elav-GAL4>UAS-Tau sham p= 0.0297. E. Quantification of latency time to copulation. For UAS-Tau sham vs. UAS-Tau injury p= 0.5777; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.0018; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.0028; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.8435; UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0074. F. Quantification of time to first wing extension by the male fly. UAS-Tau sham vs. UAS-Tau injury p= 0.2640; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p <0.0001; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.0014; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.5784; UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0045. G-N. Competitive courtship assays for UAS-Tau sham and injury male flies and Elav-GAL4>UAS-Tau sham and injury male flies (n=38, 27, 30, and 25 groups, respectively, from 10 independent trials). G. Quantification of percent of flies that mated within 2 hours. For UAS-Tau sham vs. UAS-Tau injury p= 0.1109; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.4648; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.2659; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0639. H. Quantification of latency time to copulation. For UAS-Tau sham vs. UAS-Tau injury p= 0.1535; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.0175; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.7599; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0002. I. Quantification of time to first wing extension by the male fly. For UAS-Tau sham vs. UAS-Tau injury p= 0.6975; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.0034; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.0321; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0942. J. Quantification of the total number of aggressive acts exhibited by male flies within the 10-minute window prior to copulation. For UAS-Tau sham vs. UAS-Tau injury p= 0.7363; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.3929; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.0968; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0045; UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0271. K. Quantification of the time spent engaged in aggressive acts by male flies in the 10-minute window prior to copulation. UAS-Tau sham vs. UAS-Tau injury p= 0.7200; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.3294; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.0013; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0005; UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p< 0.0001. L. Representative images of a wing pull act. Genders are indicated. Arrowheads indicate wing being pulled. M. Quantification of the total number of wing pulls by male flies in the 10-minute window prior to copulation. For UAS-Tau sham vs. UAS-Tau injury p= 0.1660; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.0434; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.0016; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0032; UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p< 0.0001. N. Quantification of the time engaged in wing pulling by male flies in the 10-minute window prior to copulation. For UAS-Tau sham vs. UAS-Tau injury p= 0.5070; UAS-Tau sham vs. Elav-GAL4>UAS-Tau sham p= 0.0753; UAS-Tau injury vs. Elav-GAL4>UAS-Tau injury p= 0.0005; Elav-GAL4>UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p= 0.0025; UAS-Tau sham vs. Elav-GAL4>UAS-Tau injury p< 0.0001. Two-sided unpaired Student’s t -tests were used to determine all p -values. For d-n graphs, all datapoints are plotted with lines indicating means ± s.d.

Article Snippet: cDNA for full-length human tau was previously purchased from Addgene (16316). cDNA for Drosophila tau was purchased from the Drosophila Genomics Resource Center (RE16764).

Techniques: Immunohistochemistry, Control

Journal: bioRxiv

Article Title: Injury-induced tau pathology promotes aggressive behavior in Drosophila without neurodegeneration

doi: 10.1101/2025.11.22.689595

Figure Lengend Snippet: Human tau expression in dopaminergic neurons coupled with injury leads to neuronal activation and increased aggression in Drosophila male flies. A. Representative max projection images of whole brain immunohistochemistry using an antibody against human tau for Ddc-GAL4, 5HT7-GAL4, Ple-GAL4, or Trh-GAL4>UAS-Tau male flies for sham and injury conditions. Scale bars: 100 μm. BC. Competitive courtship assays for sham and injury male flies expressing UASTau under Ddc-GAL4 (n=25 and 17 groups, respectively, from 8 independent trials), 5HT7-GAL4 (n=22 and 23 groups, respectively, from 7 independent trials), Ple-GAL4 (n=26 and 18 groups, respectively, from 9 independent trials), or Trh-GAL4 (n=15 and 7 groups, respectively, from 3 independent trials). Quantification for percent of flies that mated within 2 hours and latency time to copulation are in Extended Data Figure 3a-b. B. Quantification of total number of aggressive acts exhibited by male flies in the 10-minute window prior to copulation. Ddc-GAL4>UAS-Tau p= 0.2322; 5HT7-GAL4>UAS-Tau p= 0.0503; Ple-GAL4>UAS-Tau p= 0.0243; Trh-GAL4 p= 0.4566. C. Quantification of the time spent engaged in aggressive acts by male flies in the 10-minute window prior to copulation. Ddc-GAL4>UAS-Tau p= 0.0328; 5HT7-GAL4>UAS-Tau p= 0.0298; Ple-GAL4>UAS-Tau p= 0.0045; Trh-GAL4 p= 0.7751. D. Schematic of activating specific neuronal subtypes in male flies. Male flies expressing the temperature sensitive cation channel UAS-TrpA1 under Ddc-GAL4 (n=25 and 19 groups, respectively, from 4 independent trials), 5HT7-GAL4 (n=13 and 12 groups, respectively, from 2 independent trials), or Ple-GAL4 (n=26 and 15 groups, respectively, from 5 independent trials) were kept either at 26 °C to thermally activate specific neurons, or at 22 °C (no thermoactivation) for 1 hr. After 1 hr of thermal activation (or no activation), aggressive behavior was measured in a competitive courtship assay. Quantification for percent of flies that mated within 2 hours and latency time to copulation are in Extended Data Figure 3c-d. E. Quantification of total aggressive acts exhibited by male flies in the 10-minute window prior to copulation onset in a competitive courtship assay. Ddc-GAL4>UAS-TrpA1 p= 0.0044; 5HT7-GAL4>UAS-TrpA1 p= 0.9575; Ple-GAL4>UAS-TrpA1 p= 0.0084. F. Quantification of total time spent in aggressive acts by male flies in the 10-minute window prior to copulation in a competitive courtship assay. Ddc-GAL4>UAS-TrpA1 p= 0.2124; 5HT7-GAL4>UAS-TrpA1 p= 0.2470; Ple-GAL4>UAS-TrpA1 p= 0.0046. G. Representative max projection images of TUNEL staining in whole brains of UAS-Tau and Elav-GAL4>UAS-Tau male flies under sham or injury conditions. Blue outline indicates brain area. Scale bars, 100µm. H. Quantification of TUNEL positive area for each condition (n=22, 23, 38, and 36 brains). UAS-Tau sham vs. injury p= 0.1039; Elav-GAL4>UAS-Tau sham vs. injury p= 0.8935. Two-sided unpaired Student’s t -tests were used to determine p -values. All graphs display all datapoints with lines indicating means ± s.d.

Article Snippet: cDNA for full-length human tau was previously purchased from Addgene (16316). cDNA for Drosophila tau was purchased from the Drosophila Genomics Resource Center (RE16764).

Techniques: Expressing, Activation Assay, Immunohistochemistry, TUNEL Assay, Staining

Journal: bioRxiv

Article Title: Injury-induced tau pathology promotes aggressive behavior in Drosophila without neurodegeneration

doi: 10.1101/2025.11.22.689595

Figure Lengend Snippet: Human tau and fly tau compete for binding on microtubules. A-B. Competitive courtship assays for Elav-GAL4>UAS-TauS11A sham and injury flies (n=13 and 14 groups, respectively, from 3 independent trials). A. Quantification of the total number of aggressive acts exhibited by male flies within the 10-minute window prior to copulation. For Elav-GAL4>UAS-TauS11A sham vs. injury, p= 0.9406. B. Quantification of the time spent engaged in aggressive acts by male flies in the 10-minute window prior to copulation. For Elav-GAL4>UASTauS11A sham vs. injury, p= 0.9709. C. Western blot analysis of lysates generated from the heads of Elav-GAL4>UAS-Tau male flies subjected to sham or injury treatment. Blots were probed with antibodies against human tau (ms T46) and pan phospho-threonine (rb pThr). D. TIRF-M images of 2.5 nM mScarlet-2N4R-HsTau with 2.5 nM sfGFP-2N4R-HsTau show colocalization and miscibility on the microtubule, while 2.5 nM mScarlet-2N4R-HsTau and 2.5nM sfGFP-DmTau exclude each other into homotypic patches on the microtubule. Scale bars = 3 μm. E. Graph displaying individual xy pairs per pixel for mScarlet-HsTau intensity vs. sfGFP-HsTau or sfGFP-DmTau intensity on the microtubule, fit with a linear regression. Pearson’s correlation coefficients: 0.6080 for HsTau vs. HsTau (n=2042 XY pairs from n>15 microtubules, p< 0.0001) and -0.3216 for HsTau vs. DmTau (n=1830 XY pairs from n > 15 microtubules, p< 0.0001). F. TIRF-M images of 1 nM bacterially-expressed mScarlet-2N4R-HsTau in the absence or presence of 10 nM insect-cell expressed sfGFP-DmTau. Scale bars = 3 µm. G. Quantification of mScarlet-HsTau fluorescence intensity in the presence of 1 nM or 10 nM insect-cell expressed sfGFP-DmTau (n=42 and 30 microtubules, respectively from n=3 individual trials, p< 0.0001. H. TIRF-M images of 1 nM insect-cell expressed sfGFP-DmTau in the absence or presence of 10 nM bacterially-expressed mScarlet-HsTau. Scale bars = 3 µm. I. Quantification of 1 nM sfGFP-DmTau normalized fluorescence intensity in the presence of 1 nM or 10 nM bacterially-expressed mScarlet-HsTau (n=42 and 30 microtubules, respectively from n=3 individual trials, p< 0.0001). J. TIRF-M images of 1 nM insect-cell expressed sfGFP-DmTau in the presence of 20 nM or 100 nM insect-cell expressed mScarlet-HsTau. Scale bars = 3 µm. K. Quantification of 1 nM sfGFP-DmTau normalized fluorescence intensity in the presence of 1 nM, 20 nM or 100 nM insect-cell expressed mScarlet-HsTau (n=30, 47 and 33 microtubules, respectively from n=2 individual trials, **** indicates p< 0.0001). (I = insect cell expression and B = bacterial expression). Two-sided unpaired Student’s t -tests were used to determine p -values. All graphs display all datapoints with lines indicating means ± s.d.

Article Snippet: cDNA for full-length human tau was previously purchased from Addgene (16316). cDNA for Drosophila tau was purchased from the Drosophila Genomics Resource Center (RE16764).

Techniques: Binding Assay, Western Blot, Generated, Fluorescence, Expressing