Morphological phenotyping after mouse whole embryo culture.
Morphological phenotyping after mouse whole embryo culture.
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DOI：
10.3389/fcell.2023.1223849
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发表时间：
2023
影响因子：
5.5
通讯作者：
Greene, Nicholas D. E.
中科院分区：
文献类型：
--
作者：
Copp, Andrew J.;Clark, Maryam;Greene, Nicholas D. E.
小鼠胚胎的形态学表型描述在神经发育阶段，主要是作为评估胚胎8.5和9.5天之间全胚胎培养结果的指导。在此期间，神经管闭合开始并在颅区完成。在培养期结束时，脊髓闭合仍在进行中。这篇文章的重点是在表现型，可以在长凳上执行，使用立体显微镜。这包括通过卵黄囊血液循环的观察和评分来评估胚胎健康，通过some some计数来测量发育阶段，以及确定冠臀长度作为生长的衡量标准。轴向旋转（“转动”）也可以用一个简单的评分系统来评估。神经管闭合评估包括：1)确定闭合部位是否已开始闭合；2)评估颅神经形成的复杂步骤，包括从封闭点2和3开始，以及在前脑和后脑神经孔完成封闭；3)通过测量后神经孔长度评估脊柱闭合。对神经管闭合缺陷的解释需要了解，首先，特定事件预期完成的阶段，其次，胚胎标志之间的对应关系，例如，某些位置，以及由此产生的成人轴向水平。如本文所述，详细的胚胎表型与全胚胎培养的通用方法相结合，可以为早期小鼠神经发育的广泛实验研究奠定基础。
Morphological phenotyping of the mouse embryo is described at neurulation stages, primarily as a guide to evaluating the outcome of whole embryo cultures between embryonic days 8.5 and 9.5. During this period, neural tube closure is initiated and progresses to completion in the cranial region. Spinal closure is still underway at the end of the culture period. The focus of this article is particularly on phenotyping that can be performed at the bench, using a stereomicroscope. This involves assessment of embryonic health, through observation and scoring of yolk sac blood circulation, measurement of developmental stage by somite counting, and determination of crown-rump length as a measure of growth. Axial rotation (“turning”) can also be assessed using a simple scoring system. Neural tube closure assessment includes: 1) determining whether closure has been initiated at the Closure 1 site; 2) evaluating the complex steps of cranial neurulation including initiation at Closure sites 2 and 3, and completion of closure at the anterior and hindbrain neuropores; 3) assessment of spinal closure by measurement of posterior neuropore length. Interpretation of defects in neural tube closure requires an appreciation of, first, the stages that particular events are expected to be completed and, second, the correspondence between embryonic landmarks, for example, somite position, and the resulting adult axial levels. Detailed embryonic phenotyping, as described in this article, when combined with the versatile method of whole embryo culture, can form the basis for a wide range of experimental studies in early mouse neural development.
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