Analysis of mutants from a genetic screening reveals the control of intestine and liver development by many common genes in zebrafish
Analysis of mutants from a genetic screening reveals the control of intestine and liver development by many common genes in zebrafish
复制标题
对遗传筛选突变体的分析揭示了斑马鱼许多常见基因对肠道和肝脏发育的控制
DOI：
10.1016/j.bbrc.2015.03.119
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发表时间：
2015
影响因子：
3.1
通讯作者：
Huang Honghui
中科院分区：
文献类型：
--
作者：
Jiang Faming;Chen Jiehui;Ma Xirui;Huang Chao;Zhu Shicheng;Wang Fei;Li Li;Luo Lingfei;Ruan Hua;Huang Honghui
肠和肝脏都是从内胚层发育而来的，但在分子水平上，这两个消化器官在发育程序上是如何共享和不同的，我们知之甚少。一个经典的正向遗传筛选，没有基因的偏见，是一个有效的方法来解决这个问题，通过检查获得的突变体的肠和肝脏的缺陷，以评估突变的基因负责的发展，任何一个器官或两者。我们在这里报告这样的屏幕在斑马鱼。ENU具有诱变效率高、无位点选择性等优点，被用作诱变剂。在3.5dpf时收集胚胎，用肠标记fabp和肝标记fabp的混合探针进行RNA全膜原位杂交，以检查表型并确定其亲本杂合性。共鉴定了52个F2推定突变体，并且具有一般发育缺陷的突变体被流产。为了排除由高突变背景引起的不可遗传的表型，将F2推定突变体与野生型鱼异交，并在F3代中进行重新筛选。通过对来自同一F2家系的表型相似的F3突变体进行互补试验，筛选出78个诱变基因组，共鉴定出来自22个F2家系的37个F3突变系。突变体表型的分类表明，37个突变体中有31个表现出肠道和肝脏的缺陷。此外，4个肠特异性突变体和2个肝特异性突变体分别在肠和肝中表现出选择性更严重的表型。这些结果表明，肠道和肝脏在斑马鱼的两个器官的发育过程中共享大量的必需基因。对这些突变体的进一步研究将有助于深入了解斑马鱼和脊椎动物消化系统发育的分子基础。
Both the intestine and liver develop from the endoderm, yet little is known how these two digestive organs share and differ in their developmental programs, at the molecular level. A classical forward genetic screen, with no gene bias, is an effective way to address this question by examining the defects of the intestine and liver in obtained mutants to assess mutated genes responsible for the development of either organ or both. We report here such a screen in zebrafish. ENU was used as the mutagen because of its high mutagenic efficiency and no site preference. Embryos were collected at 3.5dpf for RNA whole mountin situhybridization with a cocktail probe of the intestine markerifabpand the liver markerlfabpto check phenotypes and determine their parental heterozygosis. A total of 52 F2 putative mutants were identified, and those with general developmental defects were aborted. To rule out non-inheritable phenotypes caused by high mutation background, F2 putative mutants were outcrossed with wild type fish and a re-screen in F3 generations was performed. After complementation tests between F3 mutants with similar phenotypes originating from the same F2 families, a total of 37 F3 mutant lines originated from 22 F2 families were identified after screening 78 mutagenized genomes. Classification of mutant phenotypes indicated that 31 out of the 37 mutants showed defects in both the intestine and liver. In addition, four “intestine specific mutants” and two “liver specific mutants” showed selectively more severe phenotype in the intestine and liver respectively. These results suggested that the intestine and liver share a substantial number of essential genes during both organs development in zebrafish. Further studies of the mutants are likely to shed more insights into the molecular basis of the digestive system development in the zebrafish and vertebrate.