Clinical information of this adRP family. (A) Right eye fundus of the proband (III:1) presents characteristic RP degeneration, including a waxy pallor of optic disc, attenuated artery and vein and peripheral pigment deposit. (B) OCT scan reveals macular degeneration showing reduced thickness of outer nuclear layer (ONL) and RPE at the macular area with partial loss of outer/inner segments (OS/IS). (C,D) Fundus picture and Optical coherence tomography (OCT) scans of the left eye of the unaffected member (III:3) showing as normal controls. (E) Visual fields of the right eye of the proband indicates the tubular visual field.

Genetic analyses of variants identified in the SNRNP200 Gene and its expression of SNRNP200 in cadaver eye tissues. (A) Expression of SNRNP200 in multiple cadaver eye tissues including cornea, iris, anterior capsule of lens (AC of lens), lens, posterior capsule of lens (PC of lens), ciliary body, vitreous body and neural retina and so on. retinal pigmented epithelium (RPE) is shown. A 285 bp PCR product of the SNRNP200 was detected in all tissues. A 299 bp PCR products of the human Gapdh were selected in parallel as control. (B) Sanger sequencing confirmation of the identified SNRNP200 mutations from the whole exome sequencing analysis in the recruited Chinese retinitis pigmentosa families. Sanger sequencing validation of the SNRNP200 mutations (c.6088C>T) identified by the WES analysis in this family.

The expression of SNRNP200 in zebrafish. Whole mount in situ hybridization revealed the expression of the Snrnp200 gene of zebrafish at the two-cell stage (A), 50%-epiboly (B), 8-somite stage (C), 1pdf (D), and 2pdf (E). In situ hybridization (ISH) of zebrafish retinal cryosections exhibit its expression is enriched at the ciliary marginal zone (CMZ) at 3dpf (F) and 5dpf (G).

Detrimental effects of SNRNP200 p.Arg2030Cys in zebrafish. (A) Morphological changes at 3 dpf after injections of SNRNP200^WT, SNRNP200 p.Arg2030Cys. There is a significant difference of systemic deformations among groups. (B) Statistics of normal, deformed, and dead zebrafish after injection with SNRNP200 p.Arg2030Cys, SNRNP200^WT from 2 to 4 dpf. Data are shown as mean + SD from technical triplicates. n: total number of larvae for each group from triple experiment. ***P < 0.001. (C) Immunostaining analysis of the zebrafish larvae injected with different mRNAs at 3 dpf for rhodopsin (green). The normal zebrafish revealed abundant expression of rhodopsin in the IS/OS layer (D–I) by contrast, the reactivity of rhodopsin was significantly diminished in SNRNP200^c.C6088T-injected larvae.

Phenotypic comparison among zebrafish larvae with SNRNP200 silencing, and with overexpression of SNRNP200^WT, or SNRNP200^c.C6088T. (A) Systemic phenotype of zebrafish larvae injected with 4 ng Control-MO (control); 4 ng SNRNP200-MO (MO); 4 ng SNRNP200-MO +200 pg SNRNP200^WT mRNA (MO + SNRNP200^WT); and 4 ng SNRNP200-MO+200 pg SNRNP200^c.C6088T mRNA(MO + SNRNP200^c.C6088T). (B) phenotype distribution in four groups injected variously as shown in (A) including normal, deformed and dead ones were calculated from 1 to 3 dpf. The percentage of those are shown in the graph. Comparing with the group injected with SNRNP200-MO, the group of injection of MO + SNRNP200^WT particularly reduced the ratio of deformed and dead larvae, whereas injection of MO + SNRNP200^c.C6088T significantly raise the rate of death (34 vs. 77%, P < 0.001). Data are shown as mean + SD from technical triplicates. n: total number of larvae for each group from triple experiment. **P < 0.01, ***P < 0.001.