Rod genesis driven by mafba in an nrl knockout zebrafish model with altered photoreceptor composition and progressive retinal degeneration.
Rod genesis driven by mafba in an nrl knockout zebrafish model with altered photoreceptor composition and progressive retinal degeneration.
复制标题
在具有光感受器组成改变和进行性视网膜变性的 nrl 敲除斑马鱼模型中，由 mafba 驱动的视杆发生。
DOI：
10.1371/journal.pgen.1009841
复制
发表时间：
2022-03
期刊：
影响因子：
4.5
通讯作者：
Liu M
中科院分区：
文献类型：
--
作者：
Liu F;Qin Y;Huang Y;Gao P;Li J;Yu S;Jia D;Chen X;Lv Y;Tu J;Sun K;Han Y;Reilly J;Shu X;Lu Q;Tang Z;Xu C;Luo D;Liu M
神经视网膜亮氨酸拉链（NRL）基因是决定视杆细胞命运和向视杆细胞分化的关键基因。NRL基因突变与常染色体隐性S-视锥细胞增强综合征和常染色体显性视网膜色素变性相关。然而，Nrl在调节斑马鱼（Danio rerio）（用于视网膜变性和再生研究的流行动物模型）中光感受器的发育和维持中的确切作用尚未完全确定。在这项研究中，我们通过CRISPR-Cas9技术产生了nrl敲除斑马鱼模型，并观察到令人惊讶的表型，其特征在于杆的数量减少，但不是全部损失，以及绿色视锥的过度生长。通过对斑马鱼视杆细胞发育的监测，我们发现了视杆细胞在胚胎期和胚后期的两个发育波，分别为nrl依赖型和非依赖型。通过批量和单细胞RNA测序，我们表征了来自野生型和nrl敲除斑马鱼的整个视网膜和每种视网膜细胞类型的基因表达谱。nrl突变体中绿色视锥的过度生长和视杆中绿色视锥特异性基因的错误表达表明存在视杆/绿色视锥双能前体，其在视杆与绿色视锥之间的命运选择由nrl控制。此外，基于nrl/mafba双敲除斑马鱼的细胞类型特异性表达模式和视网膜表型，我们确定mafba基因为非nrl依赖性视杆细胞发育的新调控因子。基因共线性分析揭示了mafba基因的进化起源，并提示mafba基因在竿发育中的功能是现代鱼类特有的。此外，nrl突变体中感光细胞组成的改变和异常基因表达导致进行性视网膜变性和随后的再生。因此，本研究揭示了mafba基因在斑马鱼视杆细胞发育中的新功能，并建立了斑马鱼视杆细胞和绿锥细胞光感受器发育和调控机制的工作模型。视觉是由两种类型的光传感细胞介导的，即动物眼睛中的视杆细胞和视锥细胞。感光细胞的异常生成、功能障碍或死亡都会导致不可逆的视力问题。NRL是小鼠和人类视杆细胞产生和功能的必需基因。令人惊讶的是，我们发现在人类疾病和治疗测试的流行动物模型斑马鱼中，有两种类型的视杆细胞，消除nrl基因的功能会影响胚胎期视杆细胞的形成，但不会影响幼年视杆细胞的形成。和成人阶段。胚胎后期视杆细胞的形成是由mafba基因驱动的，该基因尚未报道在视杆细胞中发挥作用。除了减少视杆细胞的数量外，nrl的缺失还导致视杆/绿色视锥杂交细胞的出现和增加的绿色视锥的数量。随后的细胞和分子改变共同导致视网膜变性。这些发现扩展了我们对光感受器发育和维持的理解，并突出了潜在的保守和物种特异性的调节机制。
Neural retina leucine zipper (NRL) is an essential gene for the fate determination and differentiation of the precursor cells into rod photoreceptors in mammals. Mutations in NRL are associated with the autosomal recessive enhanced S-cone syndrome and autosomal dominant retinitis pigmentosa. However, the exact role of Nrl in regulating the development and maintenance of photoreceptors in the zebrafish (Danio rerio), a popular animal model used for retinal degeneration and regeneration studies, has not been fully determined. In this study, we generated an nrl knockout zebrafish model via the CRISPR-Cas9 technology and observed a surprising phenotype characterized by a reduced number, but not the total loss, of rods and over-growth of green cones. We discovered two waves of rod genesis, nrl-dependent and -independent at the embryonic and post-embryonic stages, respectively, in zebrafish by monitoring the rod development. Through bulk and single-cell RNA sequencing, we characterized the gene expression profiles of the whole retina and each retinal cell type from the wild type and nrl knockout zebrafish. The over-growth of green cones and mis-expression of green-cone-specific genes in rods in nrl mutants suggested that there are rod/green-cone bipotent precursors, whose fate choice between rod versus green-cone is controlled by nrl. Besides, we identified the mafba gene as a novel regulator of the nrl-independent rod development, based on the cell-type-specific expression patterns and the retinal phenotype of nrl/mafba double-knockout zebrafish. Gene collinearity analysis revealed the evolutionary origin of mafba and suggested that the function of mafba in rod development is specific to modern fishes. Furthermore, the altered photoreceptor composition and abnormal gene expression in nrl mutants caused progressive retinal degeneration and subsequent regeneration. Accordingly, this study revealed a novel function of the mafba gene in rod development and established a working model for the developmental and regulatory mechanisms regarding the rod and green-cone photoreceptors in zebrafish. Vision is mediated by two types of light-sensing cells named rod and cone photoreceptors in animal eyes. Abnormal generation, dysfunction or death of photoreceptor cells all cause irreversible vision problems. NRL is an essential gene for the generation and function of rod cells in mice and humans. Surprisingly, we found that in the zebrafish, a popular animal model for human diseases and therapeutic testing, there are two types of rod cells, and eliminating the function of nrl gene affects the rod cell formation at the embryonic stage but not at the juvenile and adult stages. The rod cell formation at the post-embryonic is driven by the mafba gene, which has not been reported to play a role in rod cells. In addition to the reduced number of rod cells, deletion of nrl also results in the emergence of rod/green-cone hybrid cells and an increased number of green cones. The ensuing cellular and molecular alterations collectively lead to retinal degeneration. These findings expand our understanding of photoreceptor development and maintenance and highlight the underlying conserved and species-specific regulatory mechanisms.
登录
查看更多内容
影响因子：
4.5
作者：
Hao H;Kim DS;Klocke B;Johnson KR;Cui K;Gotoh N;Zang C;Gregorski J;Gieser L;Peng W;Fann Y;Seifert M;Zhao K;Swaroop A
通讯作者：
Swaroop A
影响因子：
4.5
作者：
Hanovice, Nicholas J.;Leach, Lyndsay L.;Gross, Jeffrey M.
通讯作者：
Gross, Jeffrey M.
在增强型 S 锥体综合征小鼠模型中表达视杆细胞和视锥细胞基因的混合光感受器。
影响因子：
4.5
作者：
Corbo, JC;Cepko, CL
通讯作者：
Cepko, CL
影响因子：
5.5
作者：
Angueyra JM;Kindt KS
通讯作者：
Kindt KS
影响因子：
5.3
作者：
Bernardos, Rebecca L.;Barthel, Linda K.;Raymond, Pamela A.
通讯作者：
Raymond, Pamela A.