ZFIN ID: ZDB-FIG-120412-1
Nakao et al., 2012 - The role of mislocalized phototransduction in photoreceptor cell death of retinitis pigmentosa. PLoS One   7(4):e32472 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Day 4
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage: Day 4

Fig. 1

The pathway of photoreceptor cell death does not include PDE6β.

(A) Schematic diagram of Phototransduction cascade. OS: outer segment, CC: connecting cilium, IS: inner segment, R: rhodopsin, T: transducin, PDE: phosphodiesterase (B and C) Eye ections of eyes from wt (B) and ovl (C) fish at 108 hpf. Rod photoreceptors are visualized with EGFP (Green). (Bar = 100 μm.) In ovl, the number of rod photoreceptors was decreased. (D and E) Transverse cryosections through wt (D) and ovl mutant (E) retinae at 4 dpf. F-actin is visualized with phalloidin (red), rod opsin with antibodies (green) and nuclei with Hoechst33342 (blue). Rhodopsin is mis-localized in ovl. Arrows indicate outer segments. (Bar = 10 μm.) ONL: outer nuclear layer, INL: inner nuclear layer (F and G)Ovl animals were reared in constant darkness (F) or in constant light (G) at 108 hpf. Light exposure reduces the survival of rod photoreceptor cells. (H) Graph of the number of rod photoreceptors in ovl fish at 108 hpf. (Bars mean SD, * means p<0.05.) (I) Trandsucin α expression analysis by RT-PCR of control morpholino- (lane1) and anti-Transducin α morpholino-treated (lane2). The expression of transducin is effectively suppressed by the morpholino at 108 hpf. CMO: control morpholino, TrMO: anti-Transducin α morpholino, (J and K) Sections of eyes treated by anti-transducin α morpholinos (K) and control MO (J) in ovl at 108 hpf. Anti-transducin a morpholinos rescued the rod photoreceptor cell death. (L) Rod photoreceptor numbers in anti-transducin α and control morpholino-treated ovl mutants at 108 hpf. (Bars mean SD, ** means p<0.01.) (M) PDE6β expression analysis by RT-PCR of control morpholino- (lane1) and anti-PDE6β (lane2) morpholino-treated mutants. The expression of PDE6β is effectively suppressed by the morpholino at 108 hpf. (N and O) Sections of eyes treated by anti-phosphodiesterase 6β morpholinos (O) and control MO (N) in ovl. (Bar = 100 μm.) There are no significant difference. (P) Rod photoreceptor survival in anti-phosphodiesterase 6β morpholino- (red dots) and control (black dots) morpholino-treated ovl mutants. Anti-PDE 6β morpholino has no significant effect on the number of rod photoreceptors. (Bars mean SD.)

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP ift88tz288/tz288; kj2Tg standard conditions Day 4 retinal rod cell IFL
kj2Tg control Day 4 retinal rod cell IFL
kj2Tg standard conditions Day 4 retinal rod cell IFL
kj2Tg + MO1-gnat1 standard conditions Day 4 retinal rod cell IFL
kj2Tg + MO1-pde6b standard conditions Day 4 retinal rod cell IFL
rho WT standard conditions Day 4 retinal rod cell IHC
ift88tz288/tz288 standard conditions Day 4 retinal rod cell IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ift88tz288/tz288 standard conditions Day 4 retinal rod cell pigmented, abnormal
ift88tz288/tz288 constant light Day 4 photoreceptor cell decreased amount, abnormal
ift88tz288/tz288 + MO1-gnat1 constant light Day 4 photoreceptor cell normal amount, normal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One