A technique to increase accessibility to late-stage chick embryos for in ovo manipulations.
A technique to increase accessibility to late-stage chick embryos for in ovo manipulations.
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一种增加晚期鸡胚卵内操作可及性的技术。
DOI：
10.1002/dvdy.23907
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Lwigale,Peter
中科院分区：
文献类型：
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作者：
Spurlin3rd,James;Lwigale,Peter
背景：在早期发育阶段，禽类胚胎很容易在OVO中进行移植和实验扰动。结果：在本研究中，我们描述了一种多步骤的方法，即首先在胚胎第3天开窗，然后在胚胎第5天解剖胚胎外膜，以促进胚胎在卵子中的可获得性，直到发育后期。应用该技术的胚胎大部分在E5~E8之间保持暴露，然后随着尿囊的生长从后到前逐渐移位。结论：暴露的胚胎是可存活的，并与组织移植和组织消融、电穿孔基因操作和蛋白质表达等现代发育生物学技术相兼容。这项技术为研究器官发生过程中复杂的细胞相互作用开辟了新的途径，并可以进一步推断到再生和干细胞研究。发展动力学242：148-154,2013。©2012 Wiley期刊，Inc.
Background:During early development, avian embryos are easily accessible in ovo for transplantations and experimental perturbations. However, these qualities of the avian embryonic model rapidly wane shortly after embryonic day (E)4 when the embryo is obscured by extraembryonic membranes, making it difficult to study developmental events that occur at later stages in vivo.Results:In this study, we describe a multistep method that involves initially windowing eggs at E3, followed by dissecting away extraembryonic membranes at E5 to facilitate embryo accessibility in ovo until later stages of development. The majority of the embryos subjected to this technique remain exposed between E5 and E8, then become gradually displaced by the growing allantois from posterior to anterior regions.Conclusions:Exposed embryos are viable and compatible with embryological and modern developmental biology techniques including tissue grafting and ablation, gene manipulation by electroporation, and protein expression. This technique opens up new avenues for studying complex cellular interactions during organogenesis and can be further extrapolated to regeneration and stem cell studies.Developmental Dynamics 242:148–154, 2013. © 2012 Wiley Periodicals, Inc.