An assay for lateral line regeneration in adult zebrafish.
An assay for lateral line regeneration in adult zebrafish.
复制标题
DOI：
10.3791/51343
复制
发表时间：
2014-04
期刊：
影响因子：
--
通讯作者：
Gina Pisano;Samantha M. Mason;Nyembezi Dhliwayo;R. Intine;M. Sarras
中科院分区：
文献类型：
--
作者：
Gina Pisano;Samantha M. Mason;Nyembezi Dhliwayo;R. Intine;M. Sarras
由于人类听力和平衡障碍的临床重要性，斑马鱼等模式生物已被用于研究侧线发育和再生。斑马鱼由于发育时间快，再生能力强，对这类研究特别有吸引力。到目前为止，斑马鱼侧线再生的研究主要利用胚胎和幼体阶段的鱼类，因为这些阶段的神经肥大细胞数量较少。这使得在早期发育阶段对侧线再生/或发育的定量分析变得更容易。由于许多神经系统和非神经系统疾病的斑马鱼模型是在成鱼中研究的，而不是在胚胎/仔鱼中，因此我们专注于建立一种斑马鱼成鱼侧线再生的定量检测方法，以便能够应用于当前的成鱼斑马鱼疾病模型。基于Van Trump等人之前的研究。描述了成年墨西哥盲穴鱼和斑马鱼(Danio Rerio)毛细胞的消融程序，我们的测试旨在允许在对照组和实验组之间进行定量比较。这是通过开发再生神经肥大标准曲线来实现的，该标准曲线基于庆大霉素导致侧线特定区域的毛细胞坏死后24小时内神经肥大再现的百分比。当需要更高水平的分辨率时，该分析还被设计成允许将分析扩展到单个毛细胞水平。
Due to the clinical importance of hearing and balance disorders in man, model organisms such as the zebrafish have been used to study lateral line development and regeneration. The zebrafish is particularly attractive for such studies because of its rapid development time and its high regenerative capacity. To date, zebrafish studies of lateral line regeneration have mainly utilized fish of the embryonic and larval stages because of the lower number of neuromasts at these stages. This has made quantitative analysis of lateral line regeneration/and or development easier in the earlier developmental stages. Because many zebrafish models of neurological and non-neurological diseases are studied in the adult fish and not in the embryo/larvae, we focused on developing a quantitative lateral line regenerative assay in adult zebrafish so that an assay was available that could be applied to current adult zebrafish disease models. Building on previous studies by Van Trump et al. that described procedures for ablation of hair cells in adult Mexican blind cave fish and zebrafish (Danio rerio), our assay was designed to allow quantitative comparison between control and experimental groups. This was accomplished by developing a regenerative neuromast standard curve based on the percent of neuromast reappearance over a 24 hr time period following gentamicin-induced necrosis of hair cells in a defined region of the lateral line. The assay was also designed to allow extension of the analysis to the individual hair cell level when a higher level of resolution is required.