Genetic manipulation of circadian rhythms in Xenopus.
Genetic manipulation of circadian rhythms in Xenopus.
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DOI：
10.1016/s0076-6879(05)93006-1
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发表时间：
2005
影响因子：
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通讯作者：
N. Hayasaka;Silvia I. LaRue;C. Green
中科院分区：
文献类型：
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作者：
N. Hayasaka;Silvia I. LaRue;C. Green
非洲爪蟾视网膜是研究昼夜节律振荡器机制的重要实验模型系统，因为光输入通路、中央振荡器机制和多个输出通路都包含在该组织内。这些视网膜即使在培养物中保持许多天后也继续表现出强有力的昼夜节律。该系统的有用性已得到进一步改善，甚至通过开发一种技术，这些动物的简单的遗传操作，这是补充扩大基因组学资源（非洲爪蟾基因组计划，微阵列等）。通过利用本文所述的非洲爪蟾转基因技术，可以在转基因后的几周内对原代转基因动物进行多种类型的分析。许多细胞类型特异性启动子和良好表征的细胞类型内的非洲爪蟾视网膜的可用性提供了使用这种方法的时钟功能的细胞特异性修改的优点;换句话说，昼夜节律系统内的不同细胞类型的贡献可以通过这些细胞的“分子解离”独立地分析。本文介绍了这种转基因技术是如何有用的，以及在计划和解释这些类型的实验时应考虑的各种因素。这些新技术的应用，时钟功能的研究提供了一个机会，快速评估基因表达和？或在完整视网膜的情况下发挥作用。
Xenopus laevis retina is an important experimental model system for the study of circadian oscillator mechanisms, as light input pathways, central oscillator mechanisms, and multiple output pathways are all contained within this tissue. These retinas continue to exhibit robust circadian rhythms even after being maintained in culture for many days. The usefulness of this system has been improved even further by the development of a technique for simple genetic manipulation of these animals, which is complemented by expanded genomics resources (Xenopus genome project, microarray, etc.). By taking advantage of the transgenic technique in Xenopus described in this article, many types of analysis can be done on the primary transgenic animals within a couple of weeks after transgenesis. The availability of many cell-type-specific promoters and well-characterized cell types within the Xenopus retina provides the advantage of cell-specific modification of clock function using this method; in other words, contributions of different cell types within the circadian system can be analyzed independently by" molecular dissociation" of these cells. This article describes both how this transgenic technique is useful and various considerations that should be taken into account when these types of experiments are planned and interpreted. Application of these new techniques to studies of clock function provide an opportunity to rapidly assess gene expression and? or function in the context of the intact retina.