Chen et al 2025c - Revision history

Tomamil: Tomamil moved page Review 545687567396 to Chen et al 2025c

2025-06-23T14:46:44Z

Tomamil moved page Review 545687567396 to Chen et al 2025c

Catherinechen: I removed all the [] in the methodology section

2025-06-01T00:36:42Z

I removed all the [] in the methodology section

Catherinechen: I adjusted all the comments left. I added to the discussion and future studies and clarified the methodology. I removed all the titles on the figures and fixed the citaitons to be in APA format.

2025-05-29T23:51:50Z

I adjusted all the comments left. I added to the discussion and future studies and clarified the methodology. I removed all the titles on the figures and fixed the citaitons to be in APA format.

Catherinechen at 23:31, 29 May 2025

2025-05-29T23:31:01Z

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Catherinechen: Catherinechen moved page Draft Chen 477408004 to Review 545687567396

2025-01-30T13:20:18Z

Catherinechen moved page Draft Chen 477408004 to Review 545687567396

Catherinechen: Created page with "

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 - For the alternative hypothesis, Sinapic acid will have a neuroprotective impact and reduce the severity of PD-associated defects in drosophila (climbing, memory, and dopamine). Additionally, long-term sinapic acid will have a more significant effect than Levodopa, while levodopa and sinapic acid will be the most effective. - As for our null hypothesis, Sinapic acid does not affect the flies and shouldn't be used as an alternative to Levodopa.
-==Drosophila melanogaster Strains [8]==
+==Drosophila melanogaster Strains ==
 Drosophila expressing UAS:mCD8-GFP under P{UAS(FRT.stop)mCD8-GFP.H}11 (stock #30125), TH-GAL4 under P{ple-GAL4.F}3 (stock #8848), and UAS-Hsap under P{UAS-Hsap\SNCA.F}5B (stock #8146) were obtained from the Bloomington Stock Center. All flies were maintained at 25 ± 1°C and 60% relative humidity, under a 12:12 light-dark cycle to ensure consistent environmental conditions across all assays.
-==Crossing and Generation of PD Model Flies [9]==
+==Crossing and Generation of PD Model Flies ==
 To generate the experimental lines, UAS-GFP was crossed with GAL4 for 24 hours, after which the adult flies were removed. This produced Drosophila expressing GFP in dopaminergic neurons, which were used as the control group. Separately, TH-GAL4 was crossed with UAS-Hsap to obtain Drosophila expressing mutant α-synuclein in dopaminergic neurons. These two resultant crosses were then mated to obtain the PD model: mutant α-synuclein Drosophila expressing GFP in dopaminergic neurons. Adult flies were sexed and transferred to new vials every seven days to maintain consistent density and nutrition.
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 The experimental treatment groups consisted of sinapic acid at 10, 20, and 30 µM, levodopa at 30 µM, and a combined treatment of sinapic acid and levodopa, each at 30 µM. Chemicals were administered through the media, and treatments were applied to both the GFP-expressing control flies and the PD model flies. The control group received only Carolina 4-24 Drosophila media. Each behavioral and physiological assay was performed at two time points: when the PD flies were 5 days old (young) and 35 days old (aged), with three biological replicates for each concentration and group. The independent variables were the concentrations and combinations of sinapic acid and levodopa, while the dependent variables were climbing ability, memory performance, and lifespan.
-==Rapid Iterative Negative Geotaxis (RING) Assay [10]==
+==Rapid Iterative Negative Geotaxis (RING) Assay ==
 The RING assay was used to measure locomotor ability. Vials containing flies were assembled into the RING apparatus and left undisturbed for 15–20 minutes to allow acclimation. A digital camera was placed 1 meter in front of the apparatus, aligned at mid-vial height. A timer was set for 3 seconds. The apparatus was tapped sharply on a table three times to knock all flies to the bottom of the vials, and the timer was started simultaneously with the third tap. After a 1-minute reset period, the procedure was repeated. This sequence was carried out for 5–6 trials per group. Images were uploaded to a computer and analyzed using an image viewer to count the number of flies that climbed past the 8 cm line.
-==Aversive Phototaxic Suppression (APS) Assay [11]==
+==Aversive Phototaxic Suppression (APS) Assay ==
 ===Preparation and Conditioning===
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-==Lifespan Assay [12]==
+==Lifespan Assay ==
 Three biologically identical groups were maintained for each condition to ensure statistical reliability. Each group consisted of 50–100 synchronized flies. Flies were transferred to new vials one day after birth and weekly thereafter. The number of surviving flies was recorded every 5 days until Day 35. Environmental conditions were kept constant throughout the assay.

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-[[Image:Draft_Chen_477408004-image6-c.png|center|444px]]
+[[File:Figure number 1.png|centre|444x444px|Figure #1: Climbing ability of young wild type Drosophila melanogaster. Data is the percent (± 1 SE) of the number of flies. SA1 = 10 μM; SA2 = 20 μM; SA3 = 30 μM; L3 = 30μM. #Significant to control (p<0.05) *Significant to PD(p<0.05). ^Significant to PD L3 + SA3.]]
-−
 Figure #1: Climbing ability of young wild-type Drosophila melanogaster. Data is the percent (± 1 SE) of the number of flies. SA1 = 10 μM; SA2 = 20 μM; SA3 = 30 μM; L3 = 30μM. #Significant to control (p<0.05) *Significant to PD(p<0.05). ^Significant to PD L3 + SA3.
-[[Image:Draft_Chen_477408004-image4-c.png|center|486px]]
+[[File:Figure number 2 .png|centre|485x485px|Figure #2: Climbing ability of old wild type Drosophila melanogaster. Data is the percent (± 1 SE) of the number of flies. #Significant to control (p<0.05) *Significant to PD(p<0.05). ^Significant to PD L3 + SA3]]
-−
 Figure #2: Climbing ability of young wild-type Drosophila melanogaster. Data is the percent (± 1 SE) of the number of flies. #Significant to control (p<0.05) *Significant to PD(p<0.05). ^Significant to PD L3 + SA3
@@ Line 86: / Line 84: @@
-[[Image:Draft_Chen_477408004-image5-c.png|center|600px]]
+[[File:FIgure number 3.png|centre|600x600px|Figure #3: Memory formation index for young drosophila (n=30) #Significant to control(p<0.05) *Significant to PD(p<0.05). ^Significant to PD L3 + SA3 . ]]
-−
 Figure #3: Memory formation index (n=30) #Significant to control(p<0.05) *Significant to
@@ Line 93: / Line 90: @@
-[[Image:Draft_Chen_477408004-image1-c.png|center|600px]]
+[[File:Figure number 4.png|centre|600x600px|Figure #4: Memory formation index for old drosophila (n=30) #Significant to control(p<0.05) *Significant to PD(p<0.05). ^Significant to PD L3 + SA3 .]]
-−
 Figure #4: Memory formation index (n=30) #Significant to control(p<0.05) *Significant to PD(p<0.05). ^Significant to PD L3 + SA3 .
@@ Line 101: / Line 97: @@
 Figures 5 and 6 show the lifespan of the Drosophila flies (n=30) as measured via the lifespan assay over time.
-[[Image:Draft_Chen_477408004-image3-c.png|600px]]
+[[File:Figure number 5.png|600x600px]]
 ''' '''
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-[[Image:Draft_Chen_477408004-image2-c.png|center|600px]]
+[[File:Figure number 6.png|centre|599x599px|Figure #6: Percent of drosophila alive  at day 35 (lifespan)(n=30). #Significant to control (p<0.05)*Significant to PD (p<0.05).]]
-−
 Figure #6: Percent of drosophila alive at day 35 (lifespan)(n=30). #Significant to control (p<0.05)*Significant to PD (p<0.05).

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