In Vivo Photocontrol of Microtubule Dynamics and Integrity, Migration and Mitosis, by the Potent GFP-Imaging-Compatible Photoswitchable Reagents SBTubA4P and SBTub2M.
In Vivo Photocontrol of Microtubule Dynamics and Integrity, Migration and Mitosis, by the Potent GFP-Imaging-Compatible Photoswitchable Reagents SBTubA4P and SBTub2M.
复制标题
通过有效与gfp兼容的可兼容的照片开关试剂SBTUBA4P和SBTUB2M，微管动力学和完整性，迁移和有丝分裂的体内感光控制。
DOI：
10.1021/jacs.2c01020
复制
发表时间：
2022-03-30
影响因子：
15
通讯作者：
Thorn-Seshold O
中科院分区：
文献类型：
--
作者：
Gao L;Meiring JCM;Varady A;Ruider IE;Heise C;Wranik M;Velasco CD;Taylor JA;Terni B;Weinert T;Standfuss J;Cabernard CC;Llobet A;Steinmetz MO;Bausch AR;Distel M;Thorn-Seshold J;Akhmanova A;Thorn-Seshold O
光开关试剂是细胞生物学高精度研究的有力工具。当这些试剂在二维（2D）细胞培养物中全局施用但局部光活化时，它们可以发挥微米和毫秒级的生物控制作用。这使它们在生物学上更相关的三维（3D）模型和体内，特别是用于研究具有固有时空复杂性的系统，如细胞骨架，具有很大的潜力。然而，由于在典型成像条件下的光开关异构化、代谢责任和有效浓度下的水溶性不足的组合，光开关试剂解决胞质蛋白质靶标的体内潜力在很大程度上仍未实现。在这里，我们优化了基于苯乙烯基苯并噻唑（SBT）支架的代谢稳定的类药物秋水仙素微管抑制剂的效力和溶解度，该支架对典型的荧光蛋白成像波长无反应，因此能够进行多通道成像研究。我们将这些试剂应用于3D类器官和组织外植体以及单蛋白和双蛋白成像实验中的经典模型生物（斑马鱼，爪蛙），其中时空局部照明使他们能够以细胞精度和二级分辨率在体内光控微管动力学，网络结构和微管依赖性过程。这些纳摩尔，在体内能够光开关试剂应该开辟了新的维度，货物运输，细胞运动，细胞分裂和发展的高精度细胞骨架研究。更广泛地说，它们的设计也可以激发类似的光学试剂，用于一系列细胞溶质蛋白质靶点，从而使体内药物药理学更接近一般实现。
Photoswitchable reagents are powerful tools for high-precision studies in cell biology. When these reagents are globally administered yet locally photoactivated in two-dimensional (2D) cell cultures, they can exert micron- and millisecond-scale biological control. This gives them great potential for use in biologically more relevant three-dimensional (3D) models and in vivo, particularly for studying systems with inherent spatiotemporal complexity, such as the cytoskeleton. However, due to a combination of photoswitch isomerization under typical imaging conditions, metabolic liabilities, and insufficient water solubility at effective concentrations, the in vivo potential of photoswitchable reagents addressing cytosolic protein targets remains largely unrealized. Here, we optimized the potency and solubility of metabolically stable, druglike colchicinoid microtubule inhibitors based on the styrylbenzothiazole (SBT) scaffold that are nonresponsive to typical fluorescent protein imaging wavelengths and so enable multichannel imaging studies. We applied these reagents both to 3D organoids and tissue explants and to classic model organisms (zebrafish, clawed frog) in one- and two-protein imaging experiments, in which spatiotemporally localized illuminations allowed them to photocontrol microtubule dynamics, network architecture, and microtubule-dependent processes in vivo with cellular precision and second-level resolution. These nanomolar, in vivo capable photoswitchable reagents should open up new dimensions for high-precision cytoskeleton research in cargo transport, cell motility, cell division, and development. More broadly, their design can also inspire similarly capable optical reagents for a range of cytosolic protein targets, thus bringing in vivo photopharmacology one step closer to general realization.
登录
查看更多内容
胶原蛋白的机械可塑性指导人类乳腺类器官的分支伸长。
影响因子：
16.6
作者：
Buchmann B;Engelbrecht LK;Fernandez P;Hutterer FP;Raich MK;Scheel CH;Bausch AR
通讯作者：
Bausch AR
DOI：
10.1038/s41578-021-00279-y
发表时间：
2021
期刊：
Nature reviews. Materials
影响因子：
--
作者：
Hofer M;Lutolf MP
通讯作者：
Lutolf MP
影响因子：
4.5
作者：
An, Yang;Chen, Chao;Wang, Zheng
通讯作者：
Wang, Zheng
影响因子：
16.6
作者：
Gao, Li;Meiring, Joyce C. M.;Heise, Constanze;Rai, Ankit;Mueller-Deku, Adrian;Akhmanova, Anna;Thorn-Seshold, Julia;Thorn-Seshold, Oliver
通讯作者：
Thorn-Seshold, Oliver
影响因子：
15
作者：
Borowiak M;Küllmer F;Gegenfurtner F;Peil S;Nasufovic V;Zahler S;Thorn-Seshold O;Trauner D;Arndt HD
通讯作者：
Arndt HD