Smiley Sheila, Nickerson Philip E, Comanita Lacrimioara, Daftarian Narsis, El-Sehemy Ahmed, Tsai En Leh Samuel, Matan-Lithwick Stuart, Yan Keqin, Thurig Sherry, Touahri Yacine, Dixit Rajiv, Aavani Tooka, De Repentingy Yves, Baker Adam, Tsilfidis Catherine, Biernaskie Jeff, Sauvé Yves, Schuurmans Carol, Kothary Rashmi, Mears Alan J, Wallace Valerie A
Ottawa Hospital Research Institute, 501 Smyth Road, Ottawa, Ontario, K1H 8L6, Canada.
Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
Sci Rep. 2016 Mar 11;6:22867. doi: 10.1038/srep22867.
We report successful retinal cone enrichment and transplantation using a novel cone-GFP reporter mouse line. Using the putative cone photoreceptor-enriched transcript Coiled-Coil Domain Containing 136 (Ccdc136) GFP-trapped allele, we monitored developmental reporter expression, facilitated the enrichment of cones, and evaluated transplanted GFP-labeled cones in wildtype and retinal degeneration mutant retinas. GFP reporter and endogenous Ccdc136 transcripts exhibit overlapping temporal and spatial expression patterns, both initiated in cone precursors of the embryonic retina and persisting to the adult stage in S and S/M opsin(+) cones as well as rod bipolar cells. The trapped allele does not affect cone function or survival in the adult mutant retina. When comparing the integration of GFP(+) embryonic cones and postnatal Nrl(-/-) 'cods' into retinas of adult wildtype and blind mice, both cell types integrated and exhibited a degree of morphological maturation that was dependent on donor age. These results demonstrate the amenability of the adult retina to cone transplantation using a novel transgenic resource that can advance therapeutic cone transplantation in models of age-related macular degeneration.
我们报告了使用一种新型的锥体细胞-绿色荧光蛋白(cone-GFP)报告基因小鼠品系成功实现视网膜锥体细胞富集和移植。利用假定的富含锥体细胞光感受器的转录本卷曲螺旋结构域包含蛋白136(Ccdc136)的绿色荧光蛋白捕获等位基因,我们监测了发育过程中的报告基因表达,促进了锥体细胞的富集,并评估了野生型和视网膜变性突变体视网膜中移植的绿色荧光蛋白标记的锥体细胞。绿色荧光蛋白报告基因和内源性Ccdc136转录本表现出重叠的时空表达模式,两者均在胚胎视网膜的锥体细胞前体中起始,并在S和S/M视蛋白阳性的锥体细胞以及视杆双极细胞中持续到成年阶段。捕获的等位基因不影响成年突变体视网膜中锥体细胞的功能或存活。当比较绿色荧光蛋白阳性的胚胎锥体细胞和出生后Nrl基因敲除的“视锥样细胞”(“cods”)整合到成年野生型和失明小鼠视网膜中的情况时,两种细胞类型均能整合并表现出一定程度的形态成熟,且这种成熟程度取决于供体细胞的年龄。这些结果表明,利用一种新型转基因资源,成年视网膜对锥体细胞移植具有可接受性,这可以推动在年龄相关性黄斑变性模型中的治疗性锥体细胞移植。