Alteration of limb and brain patterning in early mouse embryos by ultrasound-guided injection of Shh-expressing cells
Alteration of limb and brain patterning in early mouse embryos by ultrasound-guided injection of Shh-expressing cells
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DOI：
10.1016/s0925-4773(98)00090-2
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发表时间：
1998-07-01
影响因子：
2.6
通讯作者：
Turnbull, DH
中科院分区：
文献类型：
--
作者：
Liu, AM;Joyner, AL;Turnbull, DH
小鼠发育研究的一个基本限制是胚胎无法进入，因为胚胎被包裹在母体子宫中。我们展示了第一次使用超声背向散射显微镜引导注射细胞和其他药物进入早期小鼠胚胎。早在交配后9.5天（E9.5）就将细胞注射到小鼠神经管腔中，早在E10.5就将细胞注射到发育中的肢芽中。此外，将经工程改造以表达分泌因子Sonic Hedgehog（Shh）的细胞系注射到早期发育的小鼠脑或四肢中。Shh表达细胞被发现诱导Shh靶基因Patched和Hnf3 β在背侧脑中的异位表达，并改变前肢芽中的数字图案。这些结果表明，基因错误表达的研究，可以在小鼠胚胎中使用超声引导注射转染细胞或逆转录病毒。结合许多可用的小鼠突变体，该方法提供了一种新的方法，通过在突变小鼠背景中进行的功能获得研究来分析遗传相互作用。(C)1998爱思唯尔科学爱尔兰有限公司保留所有权利。
A basic limitation of the study of development in the mouse is the inaccessibility of the embryos, which are encased in the maternal uterus. We demonstrate the first use of ultrasound backscatter microscopy for guiding injections of cells and other agents into early stage mouse embryos. Cells were injected into the mouse neural tube cavity as early as 9.5 days post coitus (E9.5), and into the developing limb buds as early as E10.5. Furthermore, a cell line engineered to express the secreted factor Sonic Hedgehog (Shh) was injected into early developing mouse brains or limbs. The Shh-expressing cells were found to induce ectopic expression of the Shh target genes Patched and Hnf3 beta in the dorsal brain, and to alter digit patterning in the anterior limb bud. These results show that gene misexpression studies can be performed in mouse embryos using ultrasound-guided injection of transfected cells or retroviruses. In combination with the many available mouse mutants, this method offers a new approach for analyzing genetic interactions through gain-of-function studies performed in mutant mouse backgrounds. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved.