Department of Biological and Biomedical Sciences, Glasgow Caledonian University, United Kingdom.
Invest Ophthalmol Vis Sci. 2011 May 5;52(6):2960-6. doi: 10.1167/iovs.10-6800.
The authors investigated the expression and function of the zebrafish ortholog of the retinitis pigmentosa 2 (RP2) gene.
Zebrafish RP2 (ZFRP2) cDNA was isolated from adult eye mRNA by reverse transcription-polymerase chain reaction (RT-PCR). Gene expression was examined by RT-PCR. The deduced peptide sequence was aligned with RP2 orthologues from different species. Translational suppression (knockdown) of zebrafish RP2 was carried out by antisense morpholino-injection. The phenotype of ZFRP2 knockdown morphants was characterized by immunohistology and histology. Human wild-type and mutant RP2 mRNAs were coinjected with ZFRP2 morpholinos to test whether human RP2 mRNA could rescue ZFRP2 knockdown phenotypes.
ZFRP2 encodes a protein of 376 amino acids containing an N-terminal tubulin folding cofactor C-like domain and a C-terminal nucleoside diphosphate kinase-like domain. It shares 63% to 65% amino acid identity with human, mouse and bovine RP2. RP2 is expressed at the earliest stages of zebrafish development and persists into adulthood. Knockdown of RP2 in zebrafish causes a curved body axis and small eye phenotype, associated with increased cell death throughout the retina. Human wild-type RP2 mRNA could rescue the body curvature phenotype of ZFRP2 morphants, and the eye size of the resultant morphants was significantly increased over that of morphants in which ZFRP2 had been depleted.
Zebrafish RP2 is widely expressed throughout development. ZFRP2 knockdown caused retinal degeneration in zebrafish. Human RP2 could partially rescue the small eye phenotype of ZFRP2 morphants.
作者研究了斑马鱼与色素性视网膜炎 2 型（RP2）基因同源的基因的表达和功能。
通过反转录-聚合酶链反应（RT-PCR）从成年鱼眼 mRNA 中分离出斑马鱼 RP2（ZFRP2）cDNA。通过 RT-PCR 检查基因表达。将推导的肽序列与来自不同物种的 RP2 同源物进行比对。通过反义形态发生素注射进行斑马鱼 RP2 的翻译抑制（敲低）。通过免疫组织化学和组织学对 ZFRP2 敲低形态发生体的表型进行特征描述。将人野生型和突变型 RP2 mRNA 与 ZFRP2 形态发生素共注射，以测试人 RP2 mRNA 是否可以挽救 ZFRP2 敲低表型。
ZFRP2 编码一个 376 个氨基酸的蛋白质，包含一个 N 端微管折叠辅助因子 C 样结构域和一个 C 端核苷二磷酸激酶样结构域。它与人、鼠和牛 RP2 的氨基酸同一性为 63%至 65%。RP2 在斑马鱼发育的最早阶段表达，并持续到成年期。斑马鱼中 RP2 的敲低导致身体轴弯曲和眼睛小的表型，与整个视网膜细胞死亡增加有关。人野生型 RP2 mRNA 可以挽救 ZFRP2 形态发生体的身体弯曲表型，并且所得形态发生体的眼睛大小明显大于 ZFRP2 耗尽的形态发生体。
斑马鱼 RP2 在整个发育过程中广泛表达。ZFRP2 敲低导致斑马鱼视网膜变性。人 RP2 可以部分挽救 ZFRP2 形态发生体的小眼睛表型。
