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、最经典的问题之一。在许多组织中，细胞命运是由形态因子信号蛋白决定的。两个形态因子家族，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。我们和其他人已经证明，这些形态因子的分布可以通过膜突起——也被称为细胞素——来促进。细胞素很薄，富含肌动蛋白，并在细胞间运输Shh和Wnt信号通路的重要成分（Zhang和Scholpp, 2019; Curr Opin Gen & Dev）。细胞素是高度动态的，可以在几分钟内形成和收缩（Stanganello等人，2015;Nat Comms）。它们的出现受到产生细胞素的细胞及其所穿越的细胞外空间的精确控制（brent等，2021,Nat Comms）。尽管有强有力的证据表明Wnt3a和Shh在伤口愈合过程中合作，但迄今为止，尚不清楚它们的串音是如何被调节的。此外，细胞素介导的转运在伤口愈合中的作用尚不清楚。在主要细胞和发育生物学家的指导下，学生将研究Wnt和Shh在斑马鱼再生鳍中的运输。该学生将生成转基因斑马鱼系，其中信号成分被荧光标记，通过定量成像研究基于细胞素的形态素释放。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，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.