Epigenetic regulation of neuronal regeneration after spinal cord lesion in zebrafish

与哺乳动物不同，斑马鱼在受伤后可以再生脊髓神经元（Becker and Becker，2015）。这些新的神经元取代失去的神经元，并作为中继器实现功能恢复。这在哺乳动物中从未观察到。因此，重要的是要确定基因表达程序如何改变脊髓祖细胞损伤后，使神经发生在斑马鱼。在这个博士项目中，学生将阐明再生神经发生的表观遗传决定因素。Becker小组已经建立了有效的幼虫再生范例（Wehner等人，2017），初步观察表明表遗传酮乙酰化水平在病变后发生改变。目的基于这些功能数据，学生将使用麦肯齐小组（Aberdeen）的生物信息学方法来鉴定这些乙酰化改变的靶序列和下游基因（Hay et al.，2017年）。下游基因的重要性随后将通过在斑马鱼中使用CRISPR/Cas9敲除和使用TetOn系统的细胞类型特异性过表达的体内扰动实验来验证。该学生将与博士后列奥纳多卡沃内博士合作，由新的BBSRC项目资助（托马斯贝克尔，开始日期15.10.2017）。通过这项研究，我们希望阐明成功脊髓再生所需的脊髓干细胞的内在变化。该项目将为内源性哺乳动物干细胞的操作提供靶点，这些干细胞在损伤后总是无法产生神经元。

In contrast to mammals, zebrafish regenerate spinal neurons after injury (Becker and Becker, 2015).These new neurons replace lost ones and act as relays to bring about functional recovery. This is never observed in mammals. It is therefore important to determine how gene expressionprogrammes change in spinal progenitor cells after injury to allow for neurogenesis in zebrafish. In this PhD project in the Becker group (Edinburgh), the student will elucidate the epigeneticdeterminants of regenerative neurogenesis. The Becker group have established an efficient larval regeneration paradigm (Wehner et al., 2017) and preliminary observations indicate that epigenetichistone acetylation levels are altered after a lesion. Low acetylation levels are necessary for proper regeneration of motor neurons.AimsBased on these functional data the student will use bioinformatics approaches with the MacKenzie group (Aberdeen) to identify target sequences and downstream genes of these alterations ofacetylation (Hay et al., 2017). The importance of downstream genes will then be verified by in vivo perturbation experiments in zebrafish using CRISPR/Cas9 knock out and cell-type specific over- expression using the TetOn system. The student will work with a postdoc Dr Leonardo Cavone,funded by a new BBSRC project grant (to Thomas Becker, start date 15.10.2017).With this line of research, we hope to elucidate the intrinsic changes in spinal stem cells necessary for successful spinal cord regeneration. This project will provide targets for manipulations in endogenousmammalian stem cells, which invariably fail to generate neurons after injury.