Morphological analysis of cardiac malformations in developing WKY/NCrj rat embryos raised in whole embryo

全胚培养WKY/NCrj大鼠胚胎心脏畸形的形态学分析

基本信息

批准号：
05670671
负责人：
NAKAGAWA Masao
金额：
$ 1.34万
依托单位：
Shiga University of Medical Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670671/
关键词：
Whole embryo culture Sixth aortic arch HNK-1 N-CAM WKY NCrj rat Neural crest cell cardiac anomaly N-CAM

项目摘要

1.Morphological analysis of the heart in WKY/NCrj and control rat embryos raised in whole embryo culture.1) Sixth aortic arches were detected in both WKY/NCrj and control rat embryos cultured for 12 hours from 11.5 days of gestation. No morphological differences of the sixth aortic arches were observed between these two strains of embryos during 48 hours of culture.2) As the embryos cultured over 48 hours showed developmental delay compared with age-matched controls, this whole embryo culture system appeared useful only when performed for 48 hours.2.Immunolocalizations of extracellular molecules including migrating neural crest cell markers in the heart of WKY/NCrj and control rat embryos.HNK-1, N-CAM,fibronectine and cytotactin were immunopositive in the sixth aortic arches of both WKY/NCrj and control rat embryos at 11.5,12.5 and 13.5 day of gestation. The distributional patterns of those markers were almost same between WKY/NCrj and control rat embryos. N-CAM showed a chain of immunopositive tissues from nerve fibers around the foregut to the truncus arteriosus via the sixth aortic arch, which indicated that the sixth aortic arch was one of the pathways neural crest cells migrate through from splanchnic mesenchymal tissues to the truncus arteriosus. However, this study also suggested that cardiac malformations in WKY/NCrj rat might not be induced by hypoplastic sixth aortic arches or abnormal migrations of neural crest cells through this pathway.Conclusively, the hypoplastic sixth aortic arches in WKY/NCrj rat embryos at 14.5 day of gestation were supposed to be secondarily brought by an abnormal truncal division. Therefore, we are planning to clarify the distributional patterns or migratory pathways of cardiac neural crest cells which may play an important role in truncal division.

1.全胚胎培养中WKY/NCrj和对照大鼠胚胎心脏的形态学分析。1)从妊娠11.5天起培养12小时的WKY/NCrj和对照大鼠胚胎中均检测到第六主动脉弓。在48小时的培养过程中，这两个胚胎品系之间没有观察到第六主动脉弓的形态差异。2）由于培养超过48小时的胚胎与年龄匹配的对照相比显示出发育迟缓，因此整个胚胎培养系统仅在以下情况下才显得有用：进行48小时。2.细胞外分子的免疫定位，包括WKY/NCrj和对照大鼠胚胎心脏中迁移的神经嵴细胞标记。HNK-1， WKY/NCrj 和对照大鼠胚胎在妊娠 11.5、12.5 和 13.5 天的第六主动脉弓中 N-CAM、纤连蛋白和细胞趋化素均呈免疫阳性。 WKY/NCrj 和对照大鼠胚胎中这些标记的分布模式几乎相同。 N-CAM显示从前肠周围的神经纤维经第六主动脉弓到动脉干的一系列免疫阳性组织，这表明第六主动脉弓是神经嵴细胞从内脏间充质组织迁移到动脉干的途径之一。然而，这项研究还表明，WKY/NCrj 大鼠的心脏畸形可能不是由第六主动脉弓发育不良或神经嵴细胞通过该途径的异常迁移引起的。结论是，WKY/NCrj 大鼠胚胎在 14.5 天时第六主动脉弓发育不全。妊娠期的缩短应该是由异常的躯干分裂继发的。因此，我们计划阐明心脏神经嵴细胞的分布模式或迁移途径，这些细胞可能在躯干分裂中发挥重要作用。