ZFIN ID: ZDB-FIG-171211-2
Soman et al., 2017 - Inhibition of the mitochondrial calcium uniporter rescues dopaminergic neurons in pink1-/- zebrafish.. The European journal of neuroscience   45(4):528-535 Full text @ Eur. J. Neurosci.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage Range: 4-cell to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. 3 VDAC1 and MICU1 analysis. (A) RT-PCR analysis demonstrating expression of vdac1 at 1, 24, 48 and 72 hpf. (B) RT-PCR analysis of vdac1 in wt controls and after MO mediated vdac1 k/d, demonstrating a marked effect of the vdac1 MO on vdac1 wt transcript levels. (C) Dopaminergic (DA) neuronal cell count in wt, wt vdac1 (wt injected with MO against vdac1), pink1−/−, pink1−/− vdac1 (pink1−/− injected with MO against vdac1) at 3 dpf, reflecting the lack of an effect of VDAC1 inactivation on DA neuronal cell count in pink1−/− (P > 0.99) (*P = 0.0105). (D) qPCR analysis of micu1, showing significant upregulation of micu1 in pink1−/− larvae at 3 dpf compared to wt (**P = 0.0066). (E) qPCR analysis of micu2, showing a non-significant down regulation of micu2 in pink1−/− larvae at 3 dpf compared to wt (nsP = 0.0902). (F) qPCR analysis of mcur, showing a non-significant upregulation of mcur in pink1−/− larvae at 3 dpf compared to wt (nsP = 0.2088). (G) qPCR analysis of emre, showing a non-significant down regulation of emre in pink1−/− larvae at 3 dpf compared to wt (nsP = 0.2712). (H) RT-PCR analysis of micu1 at 3 dpf in uninjected larvae (WT) and after MO mediated k/d (micu MO), demonstrating the marked effect of micu1 MO injection on micu1 mRNA transcript levels. (I) Dopaminergic (DA) neuronal cell count in wt, wt micu1 (wt injected with MO against micu1), pink1/, and pink1/ micu1 (pink1−/− injected with MO against micu1) zebrafish larvae at 3 dpf, demonstrating the rescue effect of MICU1 inactivation on DA neurons in PINK1 deficiency (***P = 0.0004).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
micu1 WT standard conditions Protruding-mouth whole organism RTPCR
WT + MO1-micu1 standard conditions Protruding-mouth whole organism RTPCR
pink1sh397/sh397 standard conditions Protruding-mouth whole organism RTPCR
micu2 WT standard conditions Protruding-mouth whole organism RTPCR
pink1sh397/sh397 standard conditions Protruding-mouth whole organism RTPCR
smdt1a WT standard conditions Protruding-mouth whole organism RTPCR
pink1sh397/sh397 standard conditions Protruding-mouth whole organism RTPCR
vdac1 WT standard conditions 4-cell whole organism RTPCR
Prim-5 whole organism RTPCR
Long-pec whole organism RTPCR
Protruding-mouth whole organism RTPCR
WT + MO1-vdac1 + MO2-vdac1 standard conditions Protruding-mouth whole organism RTPCR
zgc:165532 WT standard conditions Protruding-mouth whole organism RTPCR
pink1sh397/sh397 standard conditions Protruding-mouth whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-micu1 standard conditions Protruding-mouth diencephalon has number of diencephalon dopaminergic neuron, normal
Protruding-mouth whole organism micu1 expression decreased amount, abnormal
WT + MO1-vdac1 + MO2-vdac1 standard conditions Protruding-mouth whole organism vdac1 expression decreased amount, abnormal
pink1sh397/sh397 standard conditions Protruding-mouth diencephalon has fewer parts of type diencephalon dopaminergic neuron, abnormal
pink1sh397/sh397 standard conditions Protruding-mouth whole organism micu1 expression increased amount, abnormal
pink1sh397/sh397 + MO1-micu1 standard conditions Protruding-mouth diencephalon has number of diencephalon dopaminergic neuron, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal The European journal of neuroscience for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Eur. J. Neurosci.