A transgenic zebrafish for in vivo visualization of cilia.
A transgenic zebrafish for in vivo visualization of cilia.
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作者：
纤毛是细胞信号和运动的细胞器。影响纤毛功能的突变也与纤毛相关疾病(纤毛疾病)有关。纤毛标志物的鉴定是在细胞水平上研究其功能的关键。由于缺乏一个保守的、短的纤毛定位基序，所以通常使用全长的Arl13b或5HT6蛋白来标记纤毛。然而，这些基因的过度表达会影响纤毛的功能，导致纤毛研究中的伪影。在这里，我们证明了Nephrocystin-3(NPHP3)在脊椎动物中是高度保守的，并证明了斑马鱼NPHP3的N端截短肽可以作为一个不必要的纤毛特异性标记。为了观察体内纤毛的动态变化，我们获得了稳定的转基因斑马鱼Tg(β-Actin：nphp3N-mCherry)sx1001。编码的融合蛋白可以有效地标记多种细胞类型的纤毛，从胚胎到成年期，没有任何发育和生理缺陷。我们表明，这条线允许对纤毛动态和纤毛蛋白的运输进行实时成像，例如Kif7和Smo，刺猬信号通路的关键调节因子。因此，我们已经为体内纤毛研究创造了一个有效的新工具，这将有助于进一步阐明这些重要细胞器的作用。
Cilia are organelles for cellular signalling and motility. Mutations affecting ciliary function are also associated with cilia-related disorders (ciliopathies). The identification of cilia markers is critical for studying their function at the cellular level. Due to the lack of a conserved, short ciliary localization motif, the full-length ARL13b or 5HT6 proteins are normally used for cilia labelling. Overexpression of these genes, however, can affect the function of cilia, leading to artefacts in cilia studies. Here, we show that Nephrocystin-3 (Nphp3) is highly conserved among vertebrates and demonstrate that the N-terminal truncated peptide of zebrafish Nphp3 can be used as a gratuitous cilia-specific marker. To visualize the dynamics of cilia in vivo, we generated a stable transgenic zebrafish Tg (β-actin: nphp3N-mCherry)sx1001. The cilia in multiple cell types are efficiently labelled by the encoded fusion protein from embryonic stages to adulthood, without any developmental and physiological defects. We show that the line allows live imaging of ciliary dynamics and trafficking of cilia proteins, such as Kif7 and Smo, key regulators of the Hedgehog signalling pathway. Thus, we have generated an effective new tool for in vivo cilia studies that will help shed further light on the roles of these important organelles.
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影响因子：
1.2
作者：
Ferguson, Jordan L.;Shive, Heather R.
通讯作者：
Shive, Heather R.
影响因子：
2.7
作者：
Lu, Hao;Toh, Meng Tiak;Roy, Sudipto
通讯作者：
Roy, Sudipto
影响因子：
64.5
作者：
Gerdes JM;Davis EE;Katsanis N
通讯作者：
Katsanis N
影响因子：
21.3
作者：
通讯作者：
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影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF