(A–C) Climbing activities of Ddc>G2019S flies fed on food supplemented with 0.1, 0.5, or 1.0% WGE (A), 0.1 or 1.0 mM gastrodin (B), and 0.1 or 1.0 mM 4-hydroxybenzyl alcohol (4-HBA) (C). Controls …
(A) Comparable climbing activities were detected for Ddc>mCD8-GFP and Ddc>Lrrk2 flies. Both exhibited better climbing activities than Ddc>G2019S flies in the climbing assay from weeks 1 to 6. Bar …
The climbing activities of Ddc>Lrrk2, Ddc>G2019S, and Ddc>G2019S with 0.1% WGE feeding at week 4 were assessed at weeks 3–6. Bar graphs show success rates (mean ± SEM, N = 6) of flies climbing over …
(A) Representative adult brain images showing TH-positive dopaminergic neurons in the PPL1 cluster of 2-, 4-, and 6-week-old flies of the Ddc>Lrrk2, Ddc>G2019S, and 0.1% WGE-fed Ddc>G2019S groups. …
(A–C) Representative adult whole-brain images for TH staining to reveal dopaminergic neurons in the PPL1, PPL2, PPM1/2, and PPM3 clusters of 2-, 4-, and 6-week-old flies. Scale bar: 40 μm. The …
(A) Representative immunoblots of 3-day-old adult brain lysates showing expression levels of Lrrk2, pLrrk2 (phosphorylated at Ser1292), Akt, and pAkt (phosphorylated at Ser505) in elav>Lrrk2 and elav…
(A) Representative immunoblots of 2-week-old adult brain lysates showing levels of Lrrk2 and pLrrk2 (Ser1292) in elav>Lrrk2, WGE-fed elav>Lrrk2, elav>G2019S, and WGE-fed elav>G2019S flies. (B, C) …
(A) Representative immunoblots of 2- and 4-week-old adult brain lysates showing levels of Akt and pAkt in brain extracts of elav>Lrrk2, elav>G2019S, and WGE-fed elav>G2019S flies. (B) Quantification …
(A, B) Requirement of Nrf2 in glia but not neurons for water extract of Gastrodia elata Blume (WGE)-improved Ddc>G2019S climbing activity. (A) Climbing success rates of flies in which Ddc-GAL4 …
(A, B) WGE treatment activates glial Nrf2 signals in 1-week-old ARE-GFP reporter flies. ARE-GFP (green) and glial Repo (red) in whole-mount adult brains without (A) or with 0.1% WGE treatment (B) …
WGE treatment improves the climbing ability of Ddc-LexA>G2019S flies. Bar graph shows percentage (mean ± SEM, N = 6) of flies that successfully climbed above 8 cm within 10 s. One-way ANOVA and …
(A) Nrf2 knockdown in astrocyte-like and ensheathing glia abolishes the improved locomotion elicited by WGE treatment in Ddc>G2019S flies. Composite bar graph shows climbing success rates for …
(A) Representative immunoblots of 2-week-old adult brain lysates showing protein expression levels of Lrrk2 and pLrrk2 (Ser1292) in Ddc-LexA>G2019S; repo-GAL4 and Ddc-LexA>G2019S; repo>cncC-FL2 …
(A, B) Quantifications (mean ± SEM, n > 20 for each genotype) of GFP intensities in TH-positive dopaminergic neurons nearby the astrocyte-like glia (A) or ensheathing glia (B). GFP intensities were …
(A) Representative images of glial pMad staining in the adult PPL1 clusters of Ddc>Lrrk2, Ddc>G2019S, and WGE-fed Ddc>G2019S flies. White arrows indicate pMad signals co-localized with Repo, with …
Removing one copy of Mad improves Ddc>G2019S climbing activity. Bar graphs show climbing success rates (mean ± SEM, N = 6) of 6-week-old Ddc>Lrrk2, Ddc>G2019S, and Ddc>G2019S, Mad+/− flies. One-way …
(A) Heterozygosity of Mad suppresses G2019S mutation-induced locomotion impairment in a negative geotaxis assay. Bar graph shows percentages (mean ± SEM, N = 6) of 6-week-old flies that climbed …
(A) Video-tracked paths for nTg, LRRK2-G2019S, and WGE-fed LRRK2-G2019S mice (8.5 and 11.5 months old) during the open-field test. (B) Quantification of total distance (m), average velocity (cm/s), …
(A, C) Representative images showing TH-positive dopaminergic neurons (A) or Iba-1-positive microglia (C) in the substantia nigra of 11.5-month-old nTg, LRRK2-G2019S, and WGE-fed LRRK2-G2019S mice. …
Single-channel images of Figure 10C show Iba-1-positive microglia in the substantia nigra of 11.5-month-old nTg, LRRK2-G2019S, and WGE-fed LRRK2-G2019S mice. Bars: 50 μm.
Accumulation of the hyperactivated G2019S mutant protein enhances the BMP ligand (Gbb) maturation via upregulation of Furin 1 translation in dopaminergic neurons. Secreted Gbb binds to the BMP …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (Drosophila melanogaster) | UAS-mCD8-GFP | Bloomington Drosophila Stock Center | BDSC Cat# 5137; RRID:BDSC_5137 | |
Genetic reagent (D. melanogaster) | UAS-cncTRiP | Bloomington Drosophila Stock Center | BDSC Cat# 25984; RRID:BDSC_25984 | |
Genetic reagent (D. melanogaster) | UAS-tkvQ253D | Bloomington Drosophila Stock Center | BDSC Cat# 36536; RRID:BDSC_36536 | |
Genetic reagent (D. melanogaster) | UAS-Flag- LRRK2-WT | Lin et al., 2010 | N/A | |
Genetic reagent (D. melanogaster) | UAS-Flag- LRRK2-G2019S | Lin et al., 2010 | N/A | |
Genetic reagent (D. melanogaster) | UAS-cncC-FL2 | Sykiotis and Bohmann, 2008 | N/A | |
Genetic reagent (D. melanogaster) | UAS-Mad | Takaesu et al., 2006 | N/A | |
Genetic reagent (D. melanogaster) | elav-GAL4 | Bloomington Drosophila Stock Center | BDSC Cat# 8760; RRID:BDSC_8760 | |
Genetic reagent (D. melanogaster) | repo-GAL4 | Bloomington Drosophila Stock Center | BDSC Cat# 7215; RRID:BDSC_7215 | |
Genetic reagent (D. melanogaster) | alrm-GAL4 | Bloomington Drosophila Stock Center | BDSC Cat# 67032; RRID:BDSC_67032 | |
Genetic reagent (D. melanogaster) | R56F03-GAL4 | Bloomington Drosophila Stock Center | BDSC Cat# 39157; RRID:BDSC_39157 | |
Genetic reagent (D. melanogaster) | NP2222-GAL4 | Bloomington Drosophila Stock Center | BDSC Cat# 112830; RRID:BDSC_112830 | |
Genetic reagent (D. melanogaster) | NP6293-GAL4 | Kyoto Stock Center | DGRC Cat# 105188;RRID:Kyoto Stock Center_105188 | |
Genetic reagent (D. melanogaster) | Ddc-GAL4 | Sang et al., 2007 | N/A | |
Genetic reagent (D. melanogaster) | moody-GAL4 | Bainton et al., 2005 | N/A | |
Genetic reagent (D. melanogaster) | Tub-GAL80ts | Bloomington Drosophila Stock Center | BDSC Cat# 7108; RRID:BDSC_7108 | |
Genetic reagent (D. melanogaster) | Ddc-LexA | Bloomington Drosophila Stock Center | BDSC Cat# 54218; RRID:BDSC_54218 | |
Genetic reagent (D. melanogaster) | MadK00237 | Bloomington Drosophila Stock Center | BDSC Cat# 10474; RRID:BDSC_10474 | |
Genetic reagent (D. melanogaster) | ARE-GFP | Sykiotis and Bohmann, 2008 | N/A | |
Genetic reagent (D. melanogaster) | LexAop-LRRK2-WT | This paper | N/A | See Materials and methods, ‘Drosophila stocks and maintenance’ |
Genetic reagent (D. melanogaster) | LexAop-LRRK2- G2019S | This paper | N/A | See Materials and methods, ‘Drosophila stocks and maintenance’ |
Genetic reagent (Mus musculus) | FVB/NJ | The Jackson Laboratory | JAX stock #001800 | |
Genetic reagent (M. musculus) | FVB/N-Tg(LRRK2* G2019S)1Cjli/J | The Jackson Laboratory | JAX stock #009609 | |
Antibody | Anti-TH(mouse monoclonal) | Immunostar | Cat# 22941; RRID:AB_572268 | IF (1:1000) |
Antibody | Anti-TH(rabbit polyclonal) | Millipore | Cat# AB152;RRID:AB_390204 | Mouse-IHC (1:200) |
Antibody | Anti-Repo(mouse monoclonal) | Hybridoma Bank DSHB | Cat# 8D12; RRID:AB_528448 | IF (1:500) |
Antibody | Anti-GFP(chicken polyclonal) | Abcam | Cat# ab13970; RRID:AB_300798 | IF (1:10000) |
Antibody | Anti-LRRK2(rabbit monoclonal) | Abcam | Cat# ab133474;RRID:AB_2713963 | Fly-WB (1:1000) mouse-WB(1:5000) |
Antibody | Anti-LRRK2 (phospho Ser1292)(rabbit monoclonal) | Abcam | Cat# ab203181 | Fly-WB (1:500) mouse-WB(1:1000) |
Antibody | Anti-Akt(rabbit monoclonal) | Cell Signaling | Cat# 4691;RRID:AB_915783 | WB (1:1000) |
Antibody | Anti-phospho- Drosophila Akt (Ser505)(rabbit polyclonal) | Cell Signaling | Cat# 4054;RRID:AB_331414 | WB (1:500) |
Antibody | Anti-Nrf2(rabbit polyclonal) | ThermoFisherScientific | Cat# 710574;RRID:AB_2532742 | WB (1:1000) |
Antibody | Anti-phospho- Nrf2 (Ser40)(rabbit polyclonal) | ThermoFisherScientific | Cat# PA5-67520;RRID:AB_2691678 | WB (1:1000) |
Antibody | Anti-HO-1-1(mouse monoclonal) | ThermoFisherScientific | Cat# MA1-112;RRID:AB_2536823 | WB (1:1000) |
Antibody | Anti-GAPDH(rabbit polyclonal) | GeneTex | Cat# GTX100118;RRID:AB_1080976 | WB (1:5000) |
Antibody | Anti-alpha tubulin(rabbit polyclonal) | Cell Signaling | Cat# 2144;RRID:AB_2210548 | WB (1:10,000) |
Antibody | Anti-Smad2(rabbit monoclonal) | Cell Signaling | Cat# 5339;RRID:AB_10626777 | Mouse-WB (1:1000) |
Antibody | Anti-phospho-Smad2 (Ser465/467)(rabbit monoclonal) | Cell Signaling | Cat# 3108;RRID:AB_490941 | Mouse-WB (1:1000) |
Antibody | Anti-Smad3(rabbit monoclonal) | Cell Signaling | Cat# 9523;RRID:AB_2193182 | Mouse-WB (1:1000) |
Antibody | Anti-Smad3 (phospho S423 + S425)(rabbit monoclonal) | Abcam | Cat# ab52903; RRID:AB_882596 | IF (1:250) mouse-WB (1:1000) |
Antibody | Anti-beta actin(mouse monoclonal) | Sigma-Aldrich | Cat# A5441;RRID:AB_476744 | Mouse-WB (1:5000) |
Antibody | Anti-Iba-1(rabbit polyclonal) | GeneTex | Cat# GTX100042;RRID:AB_1240434 | Mouse-IHC (1:200) |
Antibody | Anti-mouse Alexa 488(goat polyclonal) | Invitrogen | Cat# A28175;RRID:AB_2536161 | IF (1:500) |
Antibody | Anti-rabbit Alexa 488(goat polyclonal) | Invitrogen | Cat# A27034;RRID:AB_2536097 | IF (1:500) |
Antibody | Anti-rabbit DyLight 488(goat polyclonal) | ThermoFisherScientific | Cat# 35552;RRID:AB_844398 | Mouse-IHC (1:300) |
Antibody | Anti-rabbit Alexa 546(goat polyclonal) | ThermoFisherScientific | Cat# A-11035;RRID:AB_2534093 | Mouse-IHC (1:200) |
Antibody | Anti-mouse FITC(goat polyclonal) | Jackson ImmunoResearch | Cat#115-095-003;RRID:AB_2338589 | IF (1:500) |
Antibody | Anti-mouse Cy3(goat polyclonal) | Jackson ImmunoResearch | Cat# 115-165-003;RRID:AB_2338680 | IF (1:500) |
Antibody | Anti-rat IgG Cy5(goat polyclonal) | Invitrogen | Cat# A10525;RRID:AB_2534034 | IF (1:500) |
Antibody | Anti-rabbit peroxidase(goat polyclonal) | Jackson ImmunoResearch | Cat# 111-035-144;RRID:AB_2307391 | WB (1:7500) |
Antibody | Anti-mouse peroxidase (goat polyclonal) | Jackson ImmunoResearch | Cat# 115-035-003;RRID:AB_10015289 | WB (1:7500) |
Antibody | Anti-mouse peroxidase(goat polyclonal) | GeneTex | Cat# GTX213111-01;RRID:AB_10618076 | Mouse-WB (1:5000) |
Antibody | Anti-rabbit peroxidase(goat polyclonal) | GeneTex | Cat# GTX213110-01;RRID:AB_10618573 | Mouse-WB (1:5000) |
Chemical compound, drug | Phalloidin-TRITC | Sigma-Aldrich | Cat# P1951;RRID:AB_2315148 | IF (1:5000) |
Chemical compound, drug | WGE | Lin et al., 2016a;Lin et al., 2018 | N/A | KO DA Pharmaceutical Co. Ltd. |
Chemical compound, drug | Gastrodin | Lin et al., 2016b | N/A | Wuhan YC Fine Chemical Co. |
Chemical compound, drug | 4-HBA | Sigma-Aldrich | Cat# H20806-10G | |
Recombinant DNA reagent(Homo sapiens) | pDEST53-LRRK2-WT (plasmid) | Addgene | Addgene plasmid # 25044;RRID:Addgene_25044 | |
Recombinant DNA reagent(H. sapiens) | pDEST53-LRRK2-G2019S (plasmid) | Addgene | Addgene plasmid # 25045; RRID:Addgene_25045 | |
Recombinant DNA reagent(D. melanogaster) | pJFRC19-13XLexAop2-IVS- myr::GFP (plasmid) | Addgene | Addgene plasmid # 26224; RRID:Addgene_26224 | |
Software, algorithm | ImageJ | PMID:22930834 | https://imagej.nih.gov/ij;RRID:SCR_003070 | |
Software, algorithm | Ctrax | PMID:19412169 | N/A | |
Software, algorithm | CatWalk XT | Lin et al., 2020 | N/A | Noldus Information Technology |
Software, algorithm | Prism 6 | GraphPad | RRID:SCR_002798 | |
Other | DAPI | GeneTex | Cat# GTX30920 |
Statistical information.
The file includes all data and the statistical analyses in this article.
Data—western blotting.
The file includes the uncropped images of the western blotting in this article.