Yamamoto Yoshiyuki, Jeffery William R
Department of Biology, University of Maryland, College Park 20742, USA.
Methods. 2002 Dec;28(4):420-6. doi: 10.1016/s1046-2023(02)00261-x.
Experimental manipulation and other lines of evidence indicate that the lens plays a prominent role in the growth and differentiation of the vertebrate eye. Here we describe a lens transplantation method for studying the role of the lens in teleost eye development. The method involves three steps: (1) preparing embryos for the operations by embedding them in agar, (2) microsurgery with tungsten needles to remove the lens from a donor embryo and insert it into the optic cup of a host embryo lacking its own lens, and (3) a recovery period allowing surface ectoderm to close over the wound left by insertion of the lens into the host embryo. A movie illustrating the method can be found at http://www.life.umd.edu/labs/jeffery. A troubleshooting guide and summary of assays for evaluating the development of the transplanted lens and its effects on other eye parts, including the retina, are presented. Finally, some current applications of the lens transplantation method are briefly described: (1) determination of the autonomy of zebrafish lens mutants and (2) investigation of the role of the lens in eye degeneration in the cavefish Astyanax. The transplantation method will help characterize the mechanisms through which vertebrate eye development is regulated by the lens.
实验操作和其他证据表明,晶状体在脊椎动物眼睛的生长和分化中起着重要作用。在此,我们描述一种用于研究晶状体在硬骨鱼眼睛发育中作用的晶状体移植方法。该方法包括三个步骤:(1)通过将胚胎嵌入琼脂中来为手术准备胚胎;(2)用钨针进行显微手术,从供体胚胎中取出晶状体并将其插入没有自身晶状体的宿主胚胎的视杯中;(3)恢复期,使表面外胚层覆盖晶状体插入宿主胚胎后留下的伤口。可在http://www.life.umd.edu/labs/jeffery上找到说明该方法的视频。本文还提供了故障排除指南以及用于评估移植晶状体的发育及其对包括视网膜在内的其他眼部结构影响的检测方法总结。最后,简要描述了晶状体移植方法目前的一些应用:(1)确定斑马鱼晶状体突变体的自主性;(2)研究晶状体在洞穴鱼墨西哥丽脂鲤眼睛退化中的作用。该移植方法将有助于阐明晶状体调节脊椎动物眼睛发育的机制。