Adverse impact of female reproductive signaling on age-dependent neurodegeneration after mild head trauma in Drosophila
- Department of Cell Biology, Emory University School of Medicine, United States
- College of Art and Science, Emory University, United States
- Department of Neurology, Emory University School of Medicine, United States
- Center for Neurodegenerative Diseases, Emory University School of Medicine, United States
Figures
Figure 1
Exposure of adult fruit flies to a very mild form of repetitive head impacts at different ages elicits minimal acute effects.
(A) Schematic illustration of the experimental design. Male and female flies were exposed to very mild head trauma (vmHT) on D3, D17, or D31 post-eclosion (denoted as D3Inj, D17Inj, and D31Inj), …
Figure 2 with 2 supplements
Exposure to very mild head trauma (vmHT) does not acutely increase vacuole formation in the brain.
(A–C) Two-photon whole brain imaging and quantification of vacuoles in D3Inj (A), D17Inj (B), and D31Inj (C) flies. Top panels: representative z-projected whole-brain images of different injury …
Figure 2—figure supplement 1
Exposure to very mild head trauma (vmHT) on D3 does not elicit gross morphological alterations in pigment dispersing factor (PDF) neurons and mushroom bodies.
(A, B) vmHT exposure on D3 did not significantly alter PDF neuron axonal projections or dendritic arborizations when assessed 24 hr or 6 weeks after injury. (C, D) vmHT exposure on D3 did not …
Figure 2—figure supplement 2
Exposure to very mild head trauma (vmHT) does not elicit significant apoptosis.
(A–C) Representative images of TUNEL staining in Drosophila brains at 1 day post D3Inj, D17Inj, and D31Inj. There was limited TUNEL signal in both sexes and no differences in staining pattern and …
Figure 3 with 1 supplement
Exposure to very mild head trauma (vmHT) results in late-life brain deficits and neurodegeneration.
(A) Exposure to vmHT at various ages altered negative geotaxis behavior when assessed on D45. A total of nine videos were used for each condition: three experimental repeats of three trials, number …
Figure 3—figure supplement 1
Age-at-injury affects post-injury recovery after 2 weeks.
Climbing Index (CI) plots depicting sensorimotor behavior assessed two weeks following D3Inj, D17Inj, and D31Inj. Female D3Inj groups exhibited a significant decrease in negative geotaxis when …
Figure 4
AI-assisted tracking and quantification of individual fly’s behavior provides insight into defects in the speed and direction of movement.
(A) Representative snapshot of idtracker.ai-generated video at T = 6 s. Videos used for analyses were from D45 flies (see Video 1). Each tail represents a fly’s movement trajectory from the last 30 …
Figure 5 with 1 supplement
Virgin females do not exhibit neurodegenerative conditions regardless of age-at-injury.
(A) Comparison of size and mass of virgin female, mated female, and male flies. Three separate trials of n > 100 in each condition. Total mass in each trial was averaged over the number of flies to …
Figure 5—figure supplement 1
Exposure to reduced very mild head trauma (vmHT) on D3 elicited similar late-life behavioral deficits and brain pathology as exposure to regular vmHT.
(A) Climbing Index (CI) plots depicting decreases in CI in both regular D3Inj and reduced D3Inj females when climbing behavior was assessed on D45. Error bar: ±se. (B) Quantification of vacuole …
Figure 6
Eliminating SP signaling mitigates the emergence of neurodegenerative conditions in the female.
(A–C) Survival curves, modified negative geotaxis assay (mNGA) quantification, and vacuole quantification of female flies mated to SP0-males, females with pan-neuronal RNAi knockdown of SPR mated to …
Figure 7
Introducing SP signaling to pre-injured virgin females reinstates neurodegenerative phenotypes.
(A) Diagram of mating schematic and relevant behavioral and pathological assays. Virgin female flies were subjected to very mild head trauma (vmHT) on D3, whereas sham flies never received an …
Figure 8
RNA sequencing reveals sexually dimorphic and reproductive status-dependent transcriptomic responses in D3Inj fly brains.
(A) Timeline and design of RNA-seq experiment. Sham and D3Inj brains were collected at 1 day post-injury (1 dpi) and 6 weeks post-injury (6 wkpi), in four different conditions (wildtype males, …
Videos
Video 1
AI-tracked climbing behaviors of female flies.
Four groups of female flies on day 45 (D45) were subjected to the negative geotaxis assay and their climbing behaviors were recorded at a speed of 60 frames per second. From left to right, flies …
Additional files
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Supplementary file 1
Summary of the RNAseq data on transcriptomic changes and all the statistical comparison results.
(a) List of all differentially expressed genes acutely after vmHT exposure. (b) List of all differentially expressed genes at the chronic time point (day 45) after vmHT exposure on day 3. (c) List of statistical comparisons and p-values.
- https://cdn.elifesciences.org/articles/97908/elife-97908-supp1-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/97908/elife-97908-mdarchecklist1-v1.docx
