Eye Removal in Living Zebrafish Larvae to Examine Innervation-dependent Growth and Development of the Visual System

被引：1

作者：

Hagen, Olivia L. ^{[1
]}

Kim, Yehyun ^{[1
]}

Kushkowski, Elaine ^{[1
,2
]}

Rouse, Hannah ^{[3
]}

Cerveny, Kara L. ^{[1
]}

机构：

[1] Reed Coll, Dept Biol, Portland, OR 97202 USA

[2] Univ Chicago, Comm Dev Regenerat & Stem Cell Biol, Chicago, IL 60637 USA

[3] UCL, Dept Cell & Dev Biol, London, England

来源：

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2022年 / 180期

基金：

英国惠康基金;

关键词：

CELL-PROLIFERATION; OPTIC TECTUM; MICROGLIA; NERVE;

D O I：

10.3791/63509

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Zebrafish exhibit remarkable life-long growth and regenerative abilities. For example, specialized stem cell niches established during embryogenesis support continuous growth of the entire visual system, both in the eye and the brain. Coordinated growth between the retinae and the optic tectum ensures accurate retinotopic mapping as new neurons are added in the eyes and brain. To address whether retinal axons provide crucial information for regulating tectal stem and progenitor cell behaviors such as survival, proliferation, and/or differentiation, it is necessary to be able to compare innervated and denervated tectal lobes within the same animal and across animals. Surgical removal of one eye from living larval zebrafish followed by observation of the optic tectum achieves this goal. The accompanying video demonstrates how to anesthetize larvae, electrolytically sharpen tungsten needles, and use them to remove one eye. It next shows how to dissect brains from fixed zebrafish larvae. Finally, the video provides an overview of the protocol for immunohistochemistry and a demonstration of how to mount stained embryos in low-melting-point agarose for microscopy.

引用

页数：15

共 40 条

[11]

de Calbiac H., 2021, J VISUALIZED EXPT JO

[12] Imaging Subcellular Structures in the Living Zebrafish Embryo [J].