Knockdown of Laminin gamma-3 (Lamc3) impairs motoneuron guidance in the zebrafish embryo

Wellcome Open Research

article

Alexander M. J. Eve¹  James C. Smith¹ 

[1] Developmental Biology Laboratory, Francis Crick Institute

关键词: axon;    guidance;    migration;    motoneuron;    netrin;    parachordal chain;   

DOI  :  10.12688/wellcomeopenres.12394.1

学科分类：内科医学

来源: Wellcome

【 摘 要 】

Background: Previous work in the zebrafish embryo has shown that laminin γ-3 (lamc3) is enriched in endothelial cells marked by expression offli1a, but the role of Lamc3 has been unknown.Methods: We use antisense morpholino oligonucleotides, and CRISPR/Cas9 mutagenesis of F0 embryos, to create zebrafish embryos in whichlamc3 expression is compromised. Transgenic imaging, immunofluorescence, andin situ hybridisation reveal that Lamc3 loss-of-function affects the development of muscle pioneers, endothelial cells, and motoneurons.Results: Lamc3 is enriched in endothelial cells during zebrafish development, but it is also expressed by other tissues. Depletion of Lamc3 by use of antisense morpholino oligonucleotides perturbs formation of the parachordal chain and subsequently the thoracic duct, but Lamc3 is not required for sprouting of the cardinal vein. F0 embryos in whichlamc3 expression is perturbed by a CRISPR/Cas9 approach also fail to form a parachordal chain, but we were unable to establish a stablelamc3 null line. Lamc3 is dispensable for muscle pioneer specification and for the expression ofnetrin-1a in these cells. Lamc3 knockdown causesnetrin-1a up-regulation in the neural tube and there is increased Netrin-1 protein throughout the trunk of the embryo. Axonal guidance of rostral primary motoneurons is defective in Lamc3 knockdown embryos.Conclusions: We suggest that knockdown of Lamc3 perturbs migration of rostral primary motoneurons at the level of the horizontal myoseptum, indicating that laminin γ3 plays a role in motoneuron guidance.

【 授权许可】

CC BY