Fig 1.
(A) Lifespan, expressed as % survival rates, of wild type (WT) and LRRK2 flies. (B) Lifespan of untreated WT compared to treated WT, only when adults (L-/A+), with Wse, 0.1%, 1% and 10%. (C)Lifespan of untreated LRRK2 mutants compared to treated LRRK2 mutants, only when adults (L-/A+), with Withania somnifera extract (Wse), 0.1%, 1% and 10%. (D) Lifespan of untreated LRRK2 mutants compared to treated LRRK2 mutants, from their larval stage to the end of their life-cycle (L+/A+), with Wse, 0.1%, 1% and 10%. *indicates p<0.05 at Kaplan-Meier survival curves (Gehan-Breslow–Wilcoxon—Graph Pad Prism 5.01), (A) untreated LRRK2 compared to untreated WT, (B) untreated WT compared to treated WT and (C-D) untreated LRRK2 compared to treated LRRK2.
Fig 2.
Effects of Wse on climbing activity.
(A-B) Climbing activity of LRRK2 adult males treated with Wse 1% as compared with WT and untreated LRRK2 (A) and climbing activity of LRRK2 adult males treated with L-Dopa 0.01% (0.5mM) as compared with WT and untreated LRRK2 (B). Values are average ± SEM. * indicates p<0.05 at one-way ANOVA followed by LSD post hoc test as compared to WT; ** indicates p<0.05 at one-way ANOVA followed by LSD post hoc test as compared to LRRK2.
Fig 3.
Effect of LRRK2 gene mutation and treatment with Wse 1% (L-/A+) on PSP latency and amplitude recorded from Drosophila DLM.
(A) Representative traces obtained from three different flies in which PSP latency is calculated as the time (ms) from stimulus application to the peak of PSP (black arrows). (B, C) Bar graphs represent the mean ± SEM of PSP latency (ms) and amplitude (mV) recorded from flies of the indicated experimental groups. *indicates p< 0.05 compared to WT, **indicate p<0.05 compared to treated LRRK2; one-way ANOVA, followed by Bonferroni post-hoc test.
Fig 4.
Effect of LRRK2 gene mutation and treatment with Wse on the “frequency of following” recorded in Drosophila DLM.
(A) Representative traces obtained from three different flies in which PSPs were evoked in response to 10 stimulations at 100 (top) or 200 Hz (bottom). (B,C) Scatter plot graphs showing the changes in PSP amplitude following stimulation at 100 (B) or 200 Hz (C). All values are expressed as the mean ± SEM of the % relative to the amplitude of the first PSP. *indicates p< 0.05 compared to WT and Wse-untreated LRRK2 (B) and compared to WT and Wse-treated LRRK2 (C), two-way ANOVA.
Fig 5.
Samples of transmission electron microscopy images of thoracic ganglia and antennal lobes in Drosophila LRRK2 mutant (A) and after treatment with 1% in L-/A+ insects (B, C) and 10% L+/A+ (D-F) extract of Wse.
(A) abnormal mitochondria in the thoracic ganglia neuropil of Drosophila LRRK2. (B, C) conventional mitochondria in thoracic ganglia of Drosophila LRRK2 after treatment with 1% Wse L-/A+imaged at low (B) and higher magnification (C). (D, E) abnormal mitochondria in Drosophila LRRK2 thoracic ganglia cell bodies after treatment with 10% WseL+/A+. Note the irregular electron-dense substance clearly recognizable inside the mitochondria. (F and Inset) numerous endosomes are present inside the antennal lobes neurites of Drosophila LRRK2 after treatment with 10% Wse. Scale bars are 0.5 μm except in B that is 2.5 μm.