TY - JOUR
T1 - Application of local gene induction by infrared laser-mediated microscope and temperature stimulator to amphibian regeneration study
AU - Kawasumi-Kita, Aiko
AU - Hayashi, Toshinori
AU - Kobayashi, Takuya
AU - Nagayama, Chikashi
AU - Hayashi, Shinichi
AU - Kamei, Yasuhiro
AU - Morishita, Yoshihiro
AU - Takeuchi, Takashi
AU - Tamura, Koji
AU - Yokoyama, Hitoshi
N1 - Publisher Copyright:
© 2015 Japanese Society of Developmental Biologists.
PY - 2015/12/1
Y1 - 2015/12/1
N2 - Urodele amphibians (newts and salamanders) and anuran amphibians (frogs) are excellent research models to reveal mechanisms of three-dimensional organ regeneration since they have exceptionally high regenerative capacity among tetrapods. However, the difficulty in manipulating gene expression in cells in a spatially restricted manner has so far hindered elucidation of the molecular mechanisms of organ regeneration in amphibians. Recently, local heat shock by laser irradiation has enabled local gene induction even at the single-cell level in teleost fishes, nematodes, fruit flies and plants. In this study, local heat shock was made with infrared laser irradiation (IR-LEGO) by using a gene expression inducible system in transgenic animals containing a heat shock promoter, and gene expression was successfully induced only in the target region of two amphibian species, Xenopus laevis and Pleurodeles waltl (a newt), at postembryonic stages. Furthermore, we induced spatially restricted but wider gene expression in Xenopus laevis tadpoles and froglets by applying local heat shock by a temperature-controlled metal probe (temperature stimulator). The local gene manipulation systems, the IR-LEGO and the temperature stimulator, enable us to do a rigorous cell lineage trace with the combination of the Cre-LoxP system as well as to analyze gene function in a target region or cells with less off-target effects in the study of amphibian regeneration. A local heat shock by infrared laser irradiation (IR-LEGO) is applicable to amphibians to induce gene expression only at the target cells. A local heat shock by a temperature-controlled metal probe (temperature stimulator) is also applicable to an amphibian to induce wider gene expression at the target tissue. These techniques enable us to do a rigorous cell lineage trace as well as to analyze gene function in a target region or cells in study of amphibian regeneration.
AB - Urodele amphibians (newts and salamanders) and anuran amphibians (frogs) are excellent research models to reveal mechanisms of three-dimensional organ regeneration since they have exceptionally high regenerative capacity among tetrapods. However, the difficulty in manipulating gene expression in cells in a spatially restricted manner has so far hindered elucidation of the molecular mechanisms of organ regeneration in amphibians. Recently, local heat shock by laser irradiation has enabled local gene induction even at the single-cell level in teleost fishes, nematodes, fruit flies and plants. In this study, local heat shock was made with infrared laser irradiation (IR-LEGO) by using a gene expression inducible system in transgenic animals containing a heat shock promoter, and gene expression was successfully induced only in the target region of two amphibian species, Xenopus laevis and Pleurodeles waltl (a newt), at postembryonic stages. Furthermore, we induced spatially restricted but wider gene expression in Xenopus laevis tadpoles and froglets by applying local heat shock by a temperature-controlled metal probe (temperature stimulator). The local gene manipulation systems, the IR-LEGO and the temperature stimulator, enable us to do a rigorous cell lineage trace with the combination of the Cre-LoxP system as well as to analyze gene function in a target region or cells with less off-target effects in the study of amphibian regeneration. A local heat shock by infrared laser irradiation (IR-LEGO) is applicable to amphibians to induce gene expression only at the target cells. A local heat shock by a temperature-controlled metal probe (temperature stimulator) is also applicable to an amphibian to induce wider gene expression at the target tissue. These techniques enable us to do a rigorous cell lineage trace as well as to analyze gene function in a target region or cells in study of amphibian regeneration.
KW - Infrared laser-evoked gene operator
KW - Pleurodeles waltl
KW - Temperature stimulator
KW - Transgenesis
KW - Xenopus laevis
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U2 - 10.1111/dgd.12241
DO - 10.1111/dgd.12241
M3 - Article
C2 - 26510480
AN - SCOPUS:84955183900
SN - 0012-1592
VL - 57
SP - 601
EP - 613
JO - Development Growth and Differentiation
JF - Development Growth and Differentiation
IS - 9
ER -
