Dendrite Growth and Synaptogenesis in Zebrafish CNS

在活体双光子成像方法将被用来探索枝晶生长和 斑马鱼视网膜顶盖通路中的突触发生。该项目将特别关注视觉效果 刺激树突生长和突触发生的活性。斑马鱼的顶盖非常适合两个人- 光子荧光成像和视网膜顶盖投射的发展取决于活动。 新的基因编码的荧光蛋白(FP)标记和自动双光子(2P)显微镜将 用于在高时间分辨率下有效地研究树枝生长和突触发生的动力学 在很长一段时间内。本研究涉及四个具体目标：(1)确定动力学参数 在基线(黑暗，未刺激)条件下的树突生长和突触发生，(2)测量 确定的视觉刺激对树突生长和突触发生动力学参数的影响，(3)测试选择性 谷氨酸受体拮抗剂对基线和基础状态下生长和突触发生参数的影响 视觉刺激条件，以及(4)测量和分析图案化诱发的树突状钙瞬变 视觉刺激。这些研究将是第一批集成树枝乔木动态成像的研究之一 随着突触发生过程的可视化而生长。基于这一新功能的实验将测试 目前关于树突生长和突触发生动力学的各种假说相互关联， 到依赖活动的功能神经回路的发展。 要了解正常或病理性的大脑发育，显然有必要了解树突。 生长和突触发生。现在看来，树突生长和突触发生过程可能 在成熟的大脑中也同样存在，所以我们对斑马鱼发育过程的研究可能会取得进展 了解成熟神经系统的可塑性和病理学。此外，发展中的 所获得的见解可能会为L的进食性视力障碍带来意想不到的治疗机会，以及 神经创伤、神经退行性变和药物成瘾的修复作用。

In vivo two-photon imaging methods will be used to explore the mechanisms of dendrite growth and synaptogenesis in the zebrafish retinotectal pathway. The project will focus especially on effects of visually stimulated activity on dendrite growth and synaptogenesis. The zebrafish tectum is very well suited to two- photon fluorescence imaging, and development of the retinotectal projection is known to depend on activity. Novel genetically encoded fluorescent protein (FP) markers and automated two-photon (2P) microscopy will be used to efficiently study dynamics of dendritic arbor growth and synaptogenesis at high time resolution over long time periods. The research involves four specific aims: (1) determine kinetic parameters of dendrite growth and synaptogenesis under baseline (dark, unstimulated) conditions, (2) measure effects of defined visual stimuli on kinetic parameters of dendrite growth and synaptogenesis, (3) test selective glutamate receptor antagonists for effects on growth and synaptogenesis parameters under baseline and visual stimulation conditions, and (4) measure and analyze dendritic Ca transients evoked by patterned visual stimulation. These studies will be among the first to integrate dynamic imaging of dendritic arbor growth with visualization of synaptogenesis processes. Experiments based on this new capability will test various current hypotheses relating the dynamics of dendrite growth and synaptogenesis to each other and to the activity-dependent development of functional neural circuitry. To understand normal or pathological brain development, it is obviously necessary to understand dendrite growth and synaptogenesis. It now appears that dendrite growth and synaptogenesis processes may persist in mature brains, as well, so our studies of these processes in developing zebrafish may advance understanding of the plasticity and pathologies of mature nervous systems. In addition, the developmental insights obtained may point to unexpected therapeutic opportunities for l]'eating disorders of vision, and repairing effects of neurological trauma, neurodegeneration, and drug addiction.