Morphological and histological analysis of cardiovascular anomalies in congenital heart disease model rat embryos raised in whole embryo culture

全胚培养先天性心脏病模型大鼠胚胎心血管异常的形态学和组织学分析

基本信息

批准号：
07670857
负责人：
NAKAGAWA Masao
金额：
$ 1.22万
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 Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

1. Incidence of cardiovascular anomalies in bis-diamine treated rat embryos A bis-diamine administration to pregnant rats could induce cardiovascular anomalies including tetralogy of Fallot, ventral septal defect and patent truncus arteriosus. However, the incidence was different between Sprague-Dawley (SD) and Wister rats. When a single dose of 200 mg bis-diamine was administered to 9.5,10.5 and 11 pregnant day SD and Wister mother rats, the incidences of cardiovascular anomalies were 35.1%, 64.1% and 10.5%, and 100%, 92.8% and 45.6%, respectively.2. Morphological and histological analysis of the heart in Bis-diamin treated and control rat embryos raised in whole embryo culture. Single dose of 200 mg bis-diamine was administered to the pregnant Wister rats on 10.5 pregnant day. The embryos were removed at 6 hours after bis-diamine administration and immediately cultured for 24 and 48 hours following the procedures described by New et al (1973). (1) Morphological analysis : There was n … More o difinite developmental and morphological difference between bis-diamine-treated and control embryos after 24 hours culture. However, poor development of the truncal ridges could be observed in bis-diamine treated embtryos after 48 hours culture, when compared with control embyos. (2) Histological analysis : In the poorly developed truncal ridges of the bis-diamine treated embryos after 48 hours culture, there were less HNK-1 or N-CAM immunoreactive mesenchymal cells in comparison with control embryos. Furthermore, a HNK-1 or N-CAM immunoreactive chain from the neural crest to the truncus arteriosus via the 3rd, 4th and 6th aortic arches, which was clearly recognized in the control embryos after 24 or 48 hours culture, could not be detected in the bis-diamine treated ones. These findings suggested that bis-diamine prevented neural crest cells from normal migration into the heart or transformation into truncal mesenchymal cells, inducing abnormal truncal division and subsequent cardiac defects with truncal anomalies. Less

1. 双二胺处理的大鼠胚胎心血管异常的发生率怀孕大鼠给予双二胺可诱发心血管异常，包括法洛四联症、腹间隔缺损和动脉干未闭，但 Sprague-Dawley (SD) 之间的发生率不同。 ) 和 Wister 大鼠在妊娠第 9.5、10.5 和 11 天给予单剂量 200 mg 双二胺时。 Wister母鼠心血管异常发生率分别为35.1%、64.1%和10.5%，以及Bis-diamin处理和对照大鼠胚胎心脏的形态学和组织学分析。在全胚胎培养中，在怀孕 10.5 小时对怀孕的 Wister 大鼠施用单剂量 200 mg 双二胺。给予双二胺后 6 小时取出胚胎，并按照 New 等人 (1973) 描述的程序立即培养 24 和 48 小时 (1) 形态学分析：没有明确的发育和形态学分析。培养 24 小时后，双二胺处理胚胎与对照胚胎之间存在差异。然而，48 小时后，双二胺处理胚胎中可观察到躯干脊发育不良。 (2)组织学分析：培养48小时后，在双二胺处理的胚胎的发育不良的躯干脊中，与对照胚胎相比，存在较少的HNK-1或N-CAM免疫反应性间充质细胞。 N-CAM 免疫反应链从神经嵴经第 3、第 4 和第 6 动脉到达动脉干培养 24 或 48 小时后，在对照胚胎中可以清楚地识别出主动脉弓，但在双二胺处理的胚胎中却无法检测到。这些发现表明，双二胺阻止了神经嵴细胞正常迁移到心脏或转化为心脏。躯干间充质细胞，诱导异常躯干分裂和随后出现躯干异常的心脏缺陷。