Genetic analysis of the homeodomain transcription factor Chx10 in the retina using a novel multifunctional BAC transgenic mouse reporter
Genetic analysis of the homeodomain transcription factor Chx10 in the retina using a novel multifunctional BAC transgenic mouse reporter
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DOI：
10.1016/j.ydbio.2004.03.039
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发表时间：
2004-07-15
影响因子：
2.7
通讯作者：
Cepko, CL
中科院分区：
文献类型：
--
作者：
Rowan, S;Cepko, CL
Chx 10是一种含有bomeobox的转录因子，在发育中的视网膜中对祖细胞增殖和双极细胞决定至关重要。据报道，其在视网膜中的表达仅限于这些细胞群。为了进一步了解Chx 10的调控和功能，将由GFP、碱性磷酸酶和Cre重组酶组成的多功能报告基因构建体整合到编码Chx 10的BAC中。产生并分析表达该BAC的转基因小鼠的稳定品系。报告表达是忠实的内源性视网膜Chx 10的表达模式，并揭示了一个以前不受重视的位点的Chx 10表达的一个子集的Muller神经胶质细胞。此外，在成熟或（j）-Chx 10突变体视网膜中鉴定了Chx 10报告基因活性，尽管这些视网膜缺乏表达Chx 10的双极细胞。报告和分子分析表明，突变体中的表达marter的细胞具有祖细胞或部分分化的Muller神经胶质细胞的标志。这些结果强烈表明，Chx 10通过影响晚期祖细胞的分化促进双极命运。将Chx 10 BAC报告基因小鼠与R26 R小鼠杂交进行命运作图实验，结果显示，表达Chx 10转录因子的祖细胞对视网膜的所有成熟细胞类型都有贡献。这些结果证明了这些品系用于产生嵌合体或完整的视网膜遗传操作的效用。(C)2004年爱思唯尔公司All rights reserved.
Chx10 is a bomeobox-containing transcription factor critical for progenitor cell proliferation and bipolar cell determination in the developing retina. Its expression in the retina has been reported to be restricted to these cell populations. To further understand Chx10 regulation and function, a multifunctional reporter construct consisting of GFP, alkaline phosphatase, and Cre recombinase was integrated into a BAC encoding Chx10. Stable lines of transgenic mice expressing this BAC were generated and analyzed. The reporter expression was faithful to the endogenous retinal Chx10 expression pattern and revealed a previously unappreciated locus of Chx10 expression in a subset of Muller glial cells. In addition, Chx10 reporter activity was identified in mature or(j)-Chx10 mutant retinas, although these retinas lack Chx10-expressing bipolar cells. Reporter and molecular analysis showed that the reporter-expressing cells in the mutant had hallmarks of progenitor cells or partially differentiated Muller glial cells. These results strongly suggest that Chx10 promotes bipolar fate by affecting differentiation of late progenitor cells. Crosses of the Chx10 BAC reporter mice to R26R mice for fate-mapping experiments revealed that Chx10 reporter-expressing progenitor cells contribute to all mature cell types of the retina. These results demonstrate the utility of these lines for generation of mosaic or complete genetic manipulations of the retina. (C) 2004 Elsevier Inc. All rights reserved.