Regulation of Eye Regrowth in Xenopus laevis

项目总结 再生领域中的一个主要挑战是确定信号通路和合适的 组织修复所需的细胞类型。对于眼睛来说，成功的一代和 维持眼睛特异的干细胞是一个关键目标。尽管脊椎动物眼睛的发育过程 损伤后可诱导眼干细胞增殖的机制已被充分研究和表征 或者疾病仍然很难识别。这在一定程度上是因为自然干细胞的增殖通常会发生。 在胚胎发育期间，视网膜再生的研究主要利用成人(或成熟) 车型--一个非常不同的环境。高度再生的爪蛙，非洲爪蛙，是一种 建立了发育和器官再生的模型。它也与人类密切相关。我们 研究发现，非洲爪哇胚胎在5天内成功再生了功能眼。我们的研究还表明 成功的眼睛再生需要增加和延长视网膜干细胞的增殖，同时延迟 新的眼睛形态。调节眼睛再生的一个候选途径是生物电信号。生物电学 Signal在肢体再生中的作用已经得到了很好的描述，但它在眼睛发育和再生中的作用 目前还不清楚。我们的数据显示，抑制生物电信号阻止了眼睛的再生，这表明它的 函数是必需的。我们试图理解和定义生物电信号在眼睛中的作用 重新生长，作为建立一项协议的第一步，以有效剖析 开发和修复之间的区别。这个项目将：1)定义表达模式和关键 生物电信号组件在眼睛再生和发育过程中的作用；2)评估 生物电信号在激活再生视网膜前体细胞增殖和扩增中的作用 确定潜在的下游目标。总之，这项提议将利用非洲爪哇独特的生物学特性来 建立有效的策略来快速确定调节再生诱导视网膜的关键机制 体内祖细胞增殖。该协议将为最终建立一个蓝图奠定基础 富有成效的眼睛修复策略。

PROJECT SUMMARY A main challenge in the regeneration field has been to identify the signaling pathways and suitable cell types needed to enable productive tissue repair. For the eye, the successful generation and maintenance of eye-specific stem cells is a key goal. Although the process of vertebrate eye development is well-studied and characterized, the mechanisms that can induce eye stem cell proliferation following injury or disease remain difficult to identify. This is partly because natural stem cell proliferation typically occurs during embryo development whereas studies of retinal regeneration have largely utilized adult (or mature) models – a very different environment. The highly regenerative clawed frog, Xenopus laevis, is an established model for both development and organ regeneration. It is also closely related to humans. We found that Xenopus embryos successfully regrew functional eyes within 5 days. Our studies also showed that successful eye regrowth required increased and extended retinal stem cell proliferation while delaying new eye formation. One candidate pathway to regulate eye regrowth is bioelectrical signaling. Bioelectrical signaling is well-characterized for its role in limb regeneration but its role in eye development and regrowth is unclear. Our data showed that inhibition of bioelectrical signaling blocked eye regrowth, indicating that its function is required. We seek to understand and define the roles of bioelectrical signaling during eye regrowth, as a first step towards establishing a protocol for effectively dissecting the similarities and distinctions between development and repair. This project will: 1) define the expression patterns and key function of bioelectrical signaling components during eye regrowth and development; 2) assess the role of bioelectrical signaling in activating regenerative retinal progenitor cell proliferation and expansion, and 3) identify potential downstream targets. Together, this proposal will leverage the unique biology of Xenopus to establish an efficient strategy to rapidly define key mechanisms that regulate regrowth-induced retinal progenitor cell proliferation in vivo. This protocol will set the foundation for eventually building a blueprint for productive eye repair strategies.