Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers

Serizawa, Takashi; Isotani, Ayako; Matsumura, Takafumi; Nakanishi, Katsuyuki; Nonaka, Shigenori; Shibata, Shinsuke; Ikawa, Masahito; Okano, Hideyuki

doi:https://doi.org/10.1242/dev.174938

Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers

Serizawa, Takashi; Isotani, Ayako; Matsumura, Takafumi et al.

Development, 2019, 146(21), dev174938

Number of Access:883（2025-08-22 01:04 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/78565

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Title	Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers
Creator	CreatorName Serizawa, Takashi
	NRID 1000020444523 1000020444523 CreatorName Isotani, Ayako
	CreatorName Matsumura, Takafumi
	NRID 1000030263271 1000030263271 CreatorName Nakanishi, Katsuyuki
	NRID 1000090435529 1000090435529 CreatorName Nonaka, Shigenori
	NRID 1000070407089 1000070407089 CreatorName Shibata, Shinsuke
	NRID 1000020304066 1000020304066 ORCID 0000-0001-9859-6217 0000-0001-9859-6217 researchmap permalink read0054910 read0054910 Scopus Author ID 16689054500 16689054500 CreatorName Ikawa, Masahito
	NRID 1000060160694 1000060160694 CreatorName Okano, Hideyuki
Subject	2A peptide
	Fate map
	Germ layer
	Lineage-tracing system
	Site-specific recombinase
	Tissue clearing
Abstract	Genetic lineage-tracing techniques are powerful tools for studying specific cell populations in development and pathogenesis. Previous techniques have mainly involved systems for tracing a single gene, which are limited in their ability to facilitate direct comparisons of the contributions of different cell lineages. We have developed a new combinatorial system for tracing all three germ layers using self-cleaving 2A peptides and multiple site-specific recombinases (SSRs). In the resulting TRiCK (TRiple Coloured germ layer Knock-in) mice, the three germ layers are conditionally and simultaneously labelled with distinct fluorescent proteins via embryogenesis. We show that previously reported ectopic expressions of lineage markers are the outcome of secondary gene expression. The results presented here also indicate that the commitment of caudal axial stem cells to neural or mesodermal fate proceeds without lineage fluctuations, contrary to the notion of their bi-potency. Moreover, we developed IMES, an optimized tissue clearing method that is highly compatible with a variety of fluorescent proteins and immunostaining, and the combined use of TRiCK mice and IMES can facilitate comprehensive analyses of dynamic contributions of all three germ layers.
Publisher	Company of Biologists Ltd
Source Title	Development
Volume (Issue)	146(21)
Page	dev174938
Date of Issued	2019-11-04
Language	English
Handle URL	https://hdl.handle.net/11094/78565
PISSN	09501991
eISSN	14779129
NCID	AA10667805
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1242/dev.174938
Relation.isIdenticalTo	PMID 31597657
Access Rights	open access
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCTERMS.bibliographicCitation	Development. 2019, 146(21), p. dev174938
DC.title	Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers
DC.creator	Serizawa, Takashi
	Isotani, Ayako
	Matsumura, Takafumi
	Nakanishi, Katsuyuki
	Nonaka, Shigenori
	Shibata, Shinsuke
	Ikawa, Masahito
	Okano, Hideyuki
DC.publisher	Company of Biologists Ltd
DC.language' scheme='DCTERMS.RFC1766	English
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2019-11-04
DC.identifier	https://doi.org/10.1242/dev.174938
DC.subject	2A peptide
	Fate map
	Germ layer
	Lineage-tracing system
	Site-specific recombinase
	Tissue clearing
DCTERMS.abstract	Genetic lineage-tracing techniques are powerful tools for studying specific cell populations in development and pathogenesis. Previous techniques have mainly involved systems for tracing a single gene, which are limited in their ability to facilitate direct comparisons of the contributions of different cell lineages. We have developed a new combinatorial system for tracing all three germ layers using self-cleaving 2A peptides and multiple site-specific recombinases (SSRs). In the resulting TRiCK (TRiple Coloured germ layer Knock-in) mice, the three germ layers are conditionally and simultaneously labelled with distinct fluorescent proteins via embryogenesis. We show that previously reported ectopic expressions of lineage markers are the outcome of secondary gene expression. The results presented here also indicate that the commitment of caudal axial stem cells to neural or mesodermal fate proceeds without lineage fluctuations, contrary to the notion of their bi-potency. Moreover, we developed IMES, an optimized tissue clearing method that is highly compatible with a variety of fluorescent proteins and immunostaining, and the combined use of TRiCK mice and IMES can facilitate comprehensive analyses of dynamic contributions of all three germ layers.
DC.format	78565
citation_title	Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers
citation_author	Serizawa, Takashi
	Isotani, Ayako
	Matsumura, Takafumi
	Nakanishi, Katsuyuki
	Nonaka, Shigenori
	Shibata, Shinsuke
	Ikawa, Masahito
	Okano, Hideyuki
citation_publisher	Company of Biologists Ltd
citation_language	English
citation_date	2019-11-04
citation_journal_title	Development
citation_volume	146
citation_issue	21
citation_firstpage	dev174938
citation_doi	https://doi.org/10.1242/dev.174938
citation_pmid	31597657
citation_public_url	https://hdl.handle.net/11094/78565

Developmental analyses of mouse embryos and adults using a non-overlapping tracing system for all three germ layers

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