ZFIN ID: ZDB-FIG-100506-60
Sheng et al., 2010 - Deletion of the WD40 domain of LRRK2 in Zebrafish causes Parkinsonism-like loss of neurons and locomotive defect. PLoS Genetics   6(4):e1000914 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 14-19 somites to Adult
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-15 to Day 6

Fig. 3 Phenotype of neuronal loss in LRRK2 ΔWD40 morphants.

(A) Western blot analysis at 2 dpf and 3 dpf confirming the knockdown effect of zLRRK2 wild-type protein expression as well as the decreased TH protein expression in WD40 deletion morphant. Adult brain protein was used as positive control, and the ATG morphant at 3 dpf was used as negative control. The quantified results are shown in Figure S2. (B) WISH analysis at 3 dpf showing that WD40 domain deletion caused decreased TH and DAT expressions in WD40 deletion morphant, but without significant brain developmental retardation. (C) Uninjected (left) and WD40 deleted zebrafish expressing Kaede under the deltaD promoter. While neuron numbers is not much changed at 18 somite stage, there are fewer neurons in the midbrain of these 6 dpf fish, as indicated by the arrowheads. ***P<0.001, n.s.: not significant (unpaired Student′s t-test). (D) Cell apoptosis assay of uninjected (left) and WD40 delete zebrafish embryos.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
Kaede kca2Tg; rk8Tg standard conditions 14-19 somites brain neuron IFL
Day 6 brain neuron IFL
kca2Tg; rk8Tg + MO1-lrrk2 + MO2-lrrk2 standard conditions 14-19 somites brain neuron IFL
Day 6 brain neuron IFL
lrrk2 Ab1-lrrk2 WT standard conditions Long-pec whole organism WB
Ab1-lrrk2 Protruding-mouth whole organism WB
Ab1-lrrk2 Adult brain WB
Ab1-lrrk2 WT + MO1-lrrk2 + MO2-lrrk2 standard conditions Long-pec Not Detected whole organism WB
Ab1-lrrk2 Protruding-mouth Not Detected whole organism WB
Ab1-lrrk2 WT + MO3-lrrk2 standard conditions Protruding-mouth Not Detected whole organism WB
slc6a3 WT standard conditions Protruding-mouth diencephalon dopaminergic neuron ISH
WT + MO1-lrrk2 + MO2-lrrk2 standard conditions Protruding-mouth diencephalon dopaminergic neuron ISH
th WT standard conditions Long-pec whole organism WB
Protruding-mouth diencephalon dopaminergic neuron ISH
Protruding-mouth whole organism WB
WT + MO1-lrrk2 + MO2-lrrk2 standard conditions Long-pec Not Detected whole organism WB
Protruding-mouth diencephalon dopaminergic neuron ISH
Protruding-mouth Not Detected whole organism WB
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-lrrk2 + MO2-lrrk2 standard conditions Prim-15 neuron apoptotic process increased occurrence, abnormal
Protruding-mouth diencephalon dopaminergic neuron decreased amount, abnormal
kca2Tg; rk8Tg + MO1-lrrk2 + MO2-lrrk2 standard conditions Day 6 midbrain neuron decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.