Retrograde intraflagellar transport by cytoplasmic dynein-2 is required for outer segment extension in vertebrate photoreceptors but not arrestin translocation.
Retrograde intraflagellar transport by cytoplasmic dynein-2 is required for outer segment extension in vertebrate photoreceptors but not arrestin translocation.
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DOI：
10.1167/iovs.09-3828
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发表时间：
2009-11
影响因子：
4.4
通讯作者：
Perkins BD
中科院分区：
文献类型：
--
作者：
Krock BL;Mills-Henry I;Perkins BD
鞭毛内顺行运输（IFT）是光感受器外节形成和维持以及视蛋白运输所必需的。然而，逆行IFT在脊椎动物光感受器中的作用仍不清楚。本研究的目的是评估斑马鱼光感受器缺乏逆行IFT电机，细胞质动力蛋白-2。将针对细胞质动力蛋白2重链（dync 2-h1）、轻链（dync 2-li 1）和中间链（dync 2-i1）亚基的吗啉代寡核苷酸注射到斑马鱼胚胎中。用荧光化学和透射电子显微镜对这些斑马鱼变形体的视网膜和纤毛细胞进行了分析。在受精后5 - 6天（dpf）对动力蛋白变体进行全视野视网膜电图（ERG）。缺乏细胞质动力蛋白-2功能的斑马鱼表现出小眼睛、肾囊肿和短的感光细胞外节，其中一些因积累的囊泡而紊乱。Morphant感光细胞连接纤毛肿胀，但无论是视蛋白，也不arrestin是错误定位，虽然IFFT 88积累在连接纤毛的远端区域。鼻纤毛缩短，沿轴丝沿着出现胞质肿胀。细胞质动力蛋白2功能的丧失导致ERG a-、B-和d-波的振幅显著降低，但阈值反应无变化。逆行IFT对于脊椎动物光感受器的外节延伸和IFT蛋白再循环是必不可少的。结果表明，第一次，细胞质动力蛋白-2的dync 2-i1亚基是必要的逆行IFT。此外，抑制蛋白易位不需要逆行IFT。最后，ERG结果表明，细胞质动力蛋白-2的损失降低了光感受器的光反应。
Anterograde intraflagellar transport (IFT) is essential for photoreceptor outer segment formation and maintenance, as well as for opsin trafficking. However, the role of retrograde IFT in vertebrate photoreceptors remains unclear. The purpose of this study was to evaluate zebrafish photoreceptors lacking the retrograde IFT motor, cytoplasmic dynein-2. Morpholino oligonucleotides against the heavy chain (dync2-h1), light intermediate chain (dync2-li1), and intermediate chain (dync2-i1) subunits of cytoplasmic dynein-2 were injected into zebrafish embryos. Retinas and ciliated cells of these zebrafish morphants were analyzed by immunohisto-chemistry and transmission electron microscopy. Whole-field electroretinograms (ERGs) were performed on dynein morphants at 5 to 6 days after fertilization (dpf). Zebrafish lacking cytoplasmic dynein-2 function exhibited small eyes, kidney cysts, and short photoreceptor outer segments, some of which were disorganized with accumulated vesicles. Morphant photoreceptor connecting cilia were swollen, but neither opsin nor arrestin was mislocalized, although IFT88 accumulated in the distal region of the connecting cilium. Nasal cilia were shortened and displayed cytoplasmic swelling along the axoneme. Loss of cytoplasmic dynein-2 function resulted in a significant reduction in the amplitude of ERG a-, b-, and d-waves but no change in threshold response. Retrograde IFT is essential for outer segment extension and IFT protein recycling in vertebrate photoreceptors. The results show, for the first time, that the dync2-i1 subunit of cytoplasmic dynein-2 is necessary for retrograde IFT. In addition, arrestin translocation does not require retrograde IFT. Finally, the ERG results indicate that loss of cytoplasmic dynein-2 reduces the photoreceptor light response.
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影响因子：
7.8
作者：
Pazour, GJ;Wilkerson, CG;Witman, GB
通讯作者：
Witman, GB
影响因子：
16.2
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Kennedy, MJ;Dunn, FA
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Dunn, FA
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3.3
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Hou, YQ;Pazour, GJ;Witman, GB
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Witman, GB
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2.5
作者：
Insinna, Christine;Besharse, Joseph C.
通讯作者：
Besharse, Joseph C.
影响因子：
2
作者：
BROEKHUYSE, RM;TOLHUIZEN, EFJ;WINKENS, HJ
通讯作者：
WINKENS, HJ