Sox10- Venus mice: a new tool for real-time labeling of neural crest lineage cells and oligodendrocytes

Shinsuke Shibata,Akimasa Yasuda,Yukiko U Inoue,Hiroyuki Katoh,Takayoshi Inoue,Francois Renault-Mihara,Chihiro Akazawa,Momoka Sato,Hideyuki Okano,Narihito Nagoshi,Masaya Nakamura,Satoshi Suyama

doi:10.1186/1756-6606-3-31

Abstract

BackgroundWhile several mouse strains have recently been developed for tracing neural crest or oligodendrocyte lineages, each strain has inherent limitations. The connection between human SOX10 mutations and neural crest cell pathogenesis led us to focus on the Sox10 gene, which is critical for neural crest development. We generated Sox10-Venus BAC transgenic mice to monitor Sox10 expression in both normal development and in pathological processes.ResultsTissue fluorescence distinguished neural crest progeny cells and oligodendrocytes in the Sox10-Venus mouse embryo. Immunohistochemical analysis confirmed that Venus expression was restricted to cells expressing endogenous Sox10. Time-lapse imaging of various tissues in Sox10-Venus mice demonstrated that Venus expression could be visualized at the single-cell level in vivo due to the intense, focused Venus fluorescence. In the adult Sox10-Venus mouse, several types of mature and immature oligodendrocytes along with Schwann cells were clearly labeled with Venus, both before and after spinal cord injury.ConclusionsIn the newly-developed Sox10-Venus transgenic mouse, Venus fluorescence faithfully mirrors endogenous Sox10 expression and allows for in vivo imaging of live cells at the single-cell level. This Sox10-Venus mouse will thus be a useful tool for studying neural crest cells or oligodendrocytes, both in development and in pathological processes.

Highlights

While several mouse strains have recently been developed for tracing neural crest or oligodendrocyte lineages, each strain has inherent limitations
Mouse strains expressing a fluorescence-based reporter upon Cre-mediated conditional gene deletion have been developed for prospective cell sorting or direct observation without fixation [18]; the CAG-CAT-EGFP reporter transgenic mouse strain expresses enhanced green fluorescent protein (EGFP) when the loxP-flanked CAT gene located between the modified chicken b-actin promoter (CAG promoter) and the EGFP gene [18] is excised with Cre
Sox10-Venus BAC transgenic mouse generation To examine the Sox10 expression profile in vivo, we took advantage of the bacterial artificial chromosome (BAC) transgenic strategy where entire regulatory machinery for a given gene expression might be covered with a single BAC clone

Summary

Introduction

While several mouse strains have recently been developed for tracing neural crest or oligodendrocyte lineages, each strain has inherent limitations. Once a specific promoter is activated, the cell is indelibly tagged with b-galactosidase This kind of transgenic mouse is useful for monitoring the transient activation of various promoters, including the NCspecific promoter. Mouse strains expressing a fluorescence-based reporter upon Cre-mediated conditional gene deletion have been developed for prospective cell sorting or direct observation without fixation [18]; the CAG-CAT-EGFP reporter transgenic mouse strain expresses enhanced green fluorescent protein (EGFP) when the loxP-flanked CAT gene located between the modified chicken b-actin promoter (CAG promoter) and the EGFP gene [18] is excised with Cre. In previous studies, we have used mice that enable Cre/loxPmediated cell labeling with LacZ or EGFP to analyze the NC lineage and to trace NC cells after their migration and differentiation [5,19,20,21]

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Discussion

Conclusion