Optogenetic axon guidance in embryonic zebrafish

James M Harris; Andy Yu-Der Wang; Paola Arlotta

doi:10.1016/j.xpro.2021.100947

Optogenetic axon guidance in embryonic zebrafish

STAR Protoc. 2021 Nov 15;2(4):100947. doi: 10.1016/j.xpro.2021.100947. eCollection 2021 Dec 17.

Authors

James M Harris^{1

2

3}, Andy Yu-Der Wang¹, Paola Arlotta^{1

3}

Affiliations

¹ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.
² Harvard-MIT Division of Health Sciences and Technology, Harvard Medical School, Boston, MA 02115, USA.
³ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02138, USA.

Abstract

Axons form the long-range connections of biological neuronal networks, which are built through the developmental process of axon guidance. Here, we describe a protocol to precisely and non-invasively control axonal growth trajectories in live zebrafish embryos using focal light activation of a photoactivatable Rac1. We outline techniques for photostimulation, time-lapse imaging, and immunohistochemistry. These approaches enable engineering of long-range axonal circuitry or repair of defective circuits in living zebrafish, despite a milieu of competing endogenous signals and repulsive barriers. For complete details on the use and execution of this protocol, please refer to Harris et al. (2020).

Keywords: Developmental biology; Microscopy; Model Organisms; Molecular Biology; Neuroscience.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Axon Guidance* / genetics
Axon Guidance* / radiation effects
Embryo, Nonmammalian
Female
Image Processing, Computer-Assisted
Immunohistochemistry
Male
Optogenetics / methods*
Time-Lapse Imaging / methods*
Zebrafish / embryology
Zebrafish / genetics

Abstract

Publication types

MeSH terms

Grants and funding