INTRODUCTION OF A RETINOID REPORTER GENE INTO THE URODELE LIMB BLASTEMA

被引:45
作者
BROCKES, JP
机构
[1] Ludwig Institute for Cancer Research, Middlesex Hospital, University College London Branch, London W1P 8BT
关键词
D O I
10.1073/pnas.89.23.11386
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
After amputation of the limb of an adult urodele amphibian at any Point along the proximodistal axis, blastemal cells (the progenitor cells of the regenerate) give rise only to the missing structures. Retinoic acid (RA) is able to respecify the positional identity of the blastema to a more proximal value, thus raising the possibility that the RA response system is activated during limb regeneration. Cultured newt (Notophthalmus viridescens) limb cells were transfected by nuclear microinjection of plasmids which provided RA-sensitive reporter activity that could be normalized for differences in cell recovery and transfection efficiency. Such cells showed a dose-dependent response to RA in culture, and this required a functional RA response element. The cells were implanted under the wound epidermis of newt hindlimb blastemas. After injection of a proximalizing dose of RA there was a significant difference in the level of reporter activity dependent on a functional response element. When cells were implanted into contralateral proximal and distal hindlimb blastemas the proximal-to-distal ratio for activation of the reporter through the response element was approximately 3.5-fold, suggesting that a gene whose expression is regulated by RA could be differentially activated along the proximodistal axis during limb regeneration.
引用
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页码:11386 / 11390
页数:5
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