Cytoneme-mediated morphogen transport during wound healing and regeneration (ref: 4270)

细胞命运的确定是再生中最基本和最经典的问题之一。在许多组织中，细胞命运由形态发生素信号蛋白决定。形态发生素的两个家族，即Shh和Wnt，对于决定细胞增殖、分化和迁移等许多基本方面至关重要。我们和其他人已经表明，这些形态发生素的分布可以通过膜突起（也称为细胞素）来促进。细胞丝是薄的，富含肌动蛋白，并在细胞之间运输Shh和Wnt信号通路的基本组分（Zhang和Scholpp，2019; Curr Opin Gen & Dev）。细胞丝是高度动态的，并且可以在几分钟内形成和缩回（Stanganello等人，二〇一五年;它们的出现由产生细胞丝的细胞和它们穿过的细胞外空间精确控制（Brunt，et al.，2021，Nat Comms）。尽管有强有力的证据表明Wnt 3a和Shh在伤口愈合期间合作，但迄今为止，尚不清楚它们的串扰是如何调节的。此外，在伤口愈合过程中细胞丝介导的转运的作用尚不清楚。在领先的细胞和发育生物学家的监督下，学生将研究斑马鱼再生鳍中的Wnt和Shh运输。学生将生成转基因斑马鱼系，其中信号成分被荧光标记，以通过定量成像研究基于细胞丝的形态原释放。我们还将使用实时PCR、单分子荧光原位杂交和先进的高分辨率显微镜在伤口中描述信号激活。最后，学生将通过使用化学抑制剂和基于CRISPR/Cas9的突变来干扰细胞素的出现，以研究它们对斑马鱼伤口愈合的影响。

The determination of the cellular fate is one of the most fundamental and classical problems in regeneration. In many tissues, cell fate is determined by morphogen signalling proteins.Two families of morphogens, namely Shh and Wnt - are essential for determining many fundamental aspects such as cell proliferation, differentiation and migration. We and others have shown that the distribution of these morphogens can be facilitated by membrane protrusions - also known as cytonemes. Cytonemes are thin, actin-rich, and transport essential components of the Shh and Wnt signalling pathway between cells (Zhang and Scholpp, 2019; Curr Opin Gen & Dev). Cytonemes are highly dynamic and can form and retract within minutes (Stanganello et al., 2015; Nat Comms).Their emergence is precisely controlled by the cytoneme-producing cell and the extracellular space through which they traverse (Brunt, et al., 2021, Nat Comms). Although there is strong evidence that Wnt3a and Shh cooperate during wound healing, to date, it is unknown how their crosstalk is regulated. Furthermore, the role of cytoneme-mediated transport during wound healing is unclear. Under the supervision of leading cell and developmental biologists, the student will study Wnt and Shh transport in the regenerating fin in zebrafish. The student will generate transgenic zebrafish lines in which signalling components are fluorescently tagged to investigate cytoneme-based morphogen release by quantitative imaging. We will also use real-time PCR, single-molecule fluorescence in-situ hybridisation, and advanced high-resolution microscopy in the wound to describe signal activation. Finally, the student will interfere with the emergence of cytonemes by using chemical inhibitors and CRISPR/Cas9-based mutations to study their impact on zebrafish wound healing.