TY - JOUR
T1 - Fluorescent transgenic mice suitable for multi-color aggregation chimera studies
AU - Ohtsuka, Masato
AU - Miura, Hiromi
AU - Gurumurthy, Channabasavaiah B.
AU - Kimura, Minoru
AU - Inoko, Hidetoshi
AU - Yoshimura, Shinichi
AU - Sato, Masahiro
N1 - Funding Information:
Acknowledgements We thank T. Sasou, M. Ando, N. Motosugi and S. Ogiwara for technical assistance. We are grateful to Manju George for critical reading of the manuscript and for English corrections. This work was supported by Grant-in-Aid for Young Scientists (B) [20700368] from the Ministry of Education, Culture, Sports, Science and Technology (MEXT); and by the Research and Study Program of the Tokai University Educational System General Research Organization (2008) to M.O.
PY - 2012/11
Y1 - 2012/11
N2 - We recently reported a novel method of mouse transgenesis called Pronuclear Injection-based Targeted Transgenisis (PITT) using which a series of fluorescent transgenic (Tg) mice lines were generated. These lines, unlike those generated using conventional random integration methods, express the transgenes faithfully and reproducibly generation after generation. Because of this superior nature, these lines are ideal for the generation of multicolored aggregation chimeras that can be used to study cell-cell interactions and lineage analyses in living embryos/ organs, where the transgenes can be detected and the clonal origin of a given cell population easily traced by its distinct fluorescence. In this study, to verify if Tg fluorescent mice generated through PITT were suitable for such applications, we sought to generate chimeric blastocysts and chimeric-Tg mice by aggregating two- or three-colored 8- cell embryos. Our analyses using these models led to the following observations. First, we noticed that cell mixing was infrequent during the stages of morula to early blastocyst. Second, chimeric fetuses obtained after aggregation of the two-colored 8-cell embryos exhibited uniform cell mixing. And third, in the organs of adult chimeric mice, the mode of cell distribution could be either clonal or polyclonal, as previously pointed out by others. Implications of our novel and improved Tg-chimeric mice approach for clonal cell lineage and developmental studies are discussed.
AB - We recently reported a novel method of mouse transgenesis called Pronuclear Injection-based Targeted Transgenisis (PITT) using which a series of fluorescent transgenic (Tg) mice lines were generated. These lines, unlike those generated using conventional random integration methods, express the transgenes faithfully and reproducibly generation after generation. Because of this superior nature, these lines are ideal for the generation of multicolored aggregation chimeras that can be used to study cell-cell interactions and lineage analyses in living embryos/ organs, where the transgenes can be detected and the clonal origin of a given cell population easily traced by its distinct fluorescence. In this study, to verify if Tg fluorescent mice generated through PITT were suitable for such applications, we sought to generate chimeric blastocysts and chimeric-Tg mice by aggregating two- or three-colored 8- cell embryos. Our analyses using these models led to the following observations. First, we noticed that cell mixing was infrequent during the stages of morula to early blastocyst. Second, chimeric fetuses obtained after aggregation of the two-colored 8-cell embryos exhibited uniform cell mixing. And third, in the organs of adult chimeric mice, the mode of cell distribution could be either clonal or polyclonal, as previously pointed out by others. Implications of our novel and improved Tg-chimeric mice approach for clonal cell lineage and developmental studies are discussed.
KW - Aggregation chimera
KW - Clonality
KW - Fluorescent mouse
KW - Mice
KW - Preimplantation embryos
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U2 - 10.1007/s00441-012-1470-0
DO - 10.1007/s00441-012-1470-0
M3 - Article
C2 - 22868913
AN - SCOPUS:84871209001
SN - 0302-766X
VL - 350
SP - 251
EP - 260
JO - Cell and Tissue Research
JF - Cell and Tissue Research
IS - 2
ER -