Analysis of Craniocardiac Malformations in Xenopus using Optical Coherence Tomography.
Analysis of Craniocardiac Malformations in Xenopus using Optical Coherence Tomography.
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DOI：
10.1038/srep42506
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发表时间：
2017-02-14
影响因子：
4.6
通讯作者：
Khokha MK
中科院分区：
文献类型：
--
作者：
Deniz E;Jonas S;Hooper M;N Griffin J;Choma MA;Khokha MK
出生缺陷影响了美国3%的儿童。在出生缺陷中，先天性心脏病和颅面畸形是死亡和发病的主要原因。不幸的是，颅心畸形的遗传机制仍然在很大程度上不明。为了解决这个问题，人类基因组研究正在确定患者的序列变异，从而产生许多候选基因。然而，大多数候选基因的发病机制尚不清楚。因此，需要在人类疾病的快速和有效的动物模型中进行功能分析。在这里，我们耦合青蛙爪蟾热带光学相干断层扫描（OCT），以创建一个快速，高效的系统，用于测试颅心候选基因。OCT可以以接近组织学的分辨率对体内微观结构的横截面进行成像。在这里，我们确定最佳的OCT成像平面可视化和定量爪蟾心脏和面部结构建立规范的数据。接下来，我们评估已知的人类先天性心脏病：心肌病和心脏异位。最后，我们研究颅面缺陷的一个已知的人类致畸剂，环巴胺。我们很容易概括人类的表型，并量化功能和结构缺陷。使用这种方法，我们可以快速测试人类颅心候选基因的表型，作为理解候选基因的疾病机制的关键的第一步。
Birth defects affect 3% of children in the United States. Among the birth defects, congenital heart disease and craniofacial malformations are major causes of mortality and morbidity. Unfortunately, the genetic mechanisms underlying craniocardiac malformations remain largely uncharacterized. To address this, human genomic studies are identifying sequence variations in patients, resulting in numerous candidate genes. However, the molecular mechanisms of pathogenesis for most candidate genes are unknown. Therefore, there is a need for functional analyses in rapid and efficient animal models of human disease. Here, we coupled the frog Xenopus tropicalis with Optical Coherence Tomography (OCT) to create a fast and efficient system for testing craniocardiac candidate genes. OCT can image cross-sections of microscopic structures in vivo at resolutions approaching histology. Here, we identify optimal OCT imaging planes to visualize and quantitate Xenopus heart and facial structures establishing normative data. Next we evaluate known human congenital heart diseases: cardiomyopathy and heterotaxy. Finally, we examine craniofacial defects by a known human teratogen, cyclopamine. We recapitulate human phenotypes readily and quantify the functional and structural defects. Using this approach, we can quickly test human craniocardiac candidate genes for phenocopy as a critical first step towards understanding disease mechanisms of the candidate genes.