Gap junction-mediated cell-cell communication modulates mouse neural crest migration.
Gap junction-mediated cell-cell communication modulates mouse neural crest migration.
复制标题
间隙连接介导的细胞间通讯调节小鼠神经嵴迁移。
DOI：
10.1083/jcb.143.6.1725
复制
发表时间：
1998-12-14
影响因子：
7.8
通讯作者：
Lo, C W
中科院分区：
文献类型：
--
作者：
Huang, G Y;Cooper, E S;Waldo, K;Kirby, M L;Gilula, N B;Lo, C W
既往研究表明，神经嵴细胞α - 1连接蛋白功能的升高或降低可引起锥体状心脏畸形。为了阐明α1连接蛋白缺口连接在心脏发育中定量需求的可能依据，我们利用神经嵴生长培养系统检测了过表达α1连接蛋白的CMV43转基因胚胎、α1连接蛋白敲除（KO）小鼠和表达α1连接蛋白显性阴性的FC转基因小鼠的神经嵴细胞的迁移情况。这些研究表明，CMV43胚胎的心神经嵴迁移率增加，而FC转基因和α1连接蛋白KO胚胎的心神经嵴迁移率降低。纯合子α1连接蛋白KO和半合子α1连接蛋白KO和CMV43胚胎的迁移变化分别与连接蛋白基因或转基因剂量同步发生。对离体培养物和活胚中神经嵴细胞的染料偶联分析显示，CMV43转基因小鼠的间隙连接通讯增加，而FC转基因小鼠和α1连接蛋白KO小鼠的间隙连接通讯减少。进一步的分析表明，在非转基因外植体培养中，使用油酰胺下调间隙连接的通信表明，这种减少心脏嵴细胞间隙连接通信的独立方法也导致了嵴迁移速率的降低。为了确定这些发现与体内神经嵴迁移的可能相关性，研究人员使用lacZ转基因来观察心脏神经嵴细胞在流出道中的分布。这些研究表明，在CMV43转基因小鼠流出隔中有更多的lacz阳性细胞，而在α1连接蛋白KO小鼠中则有减少。令人惊讶的是，这伴随着细胞增殖的变化，而不是在心脏神经嵴细胞中，而是在心肌中——CMV43小鼠的细胞增殖升高，而α1连接蛋白KO小鼠的细胞增殖降低。后一种观察结果表明，心脏神经嵴细胞可能在调节非神经嵴来源组织的生长发育中起作用。综上所述，α1连接蛋白介导的间隙连接通讯在心脏神经嵴迁移中起重要作用。此外，它们表明心脏神经嵴扰动可能是α1连接蛋白功能获得或丧失的小鼠心脏缺陷的潜在原因。
Previous studies showed that conotruncal heart malformations can arise with the increase or decrease in α1 connexin function in neural crest cells. To elucidate the possible basis for the quantitative requirement for α1 connexin gap junctions in cardiac development, a neural crest outgrowth culture system was used to examine migration of neural crest cells derived from CMV43 transgenic embryos overexpressing α1 connexins, and from α1 connexin knockout (KO) mice and FC transgenic mice expressing a dominant-negative α1 connexin fusion protein. These studies showed that the migration rate of cardiac neural crest was increased in the CMV43 embryos, but decreased in the FC transgenic and α1 connexin KO embryos. Migration changes occurred in step with connexin gene or transgene dosage in the homozygous vs. hemizygous α1 connexin KO and CMV43 embryos, respectively. Dye coupling analysis in neural crest cells in the outgrowth cultures and also in the living embryos showed an elevation of gap junction communication in the CMV43 transgenic mice, while a reduction was observed in the FC transgenic and α1 connexin KO mice. Further analysis using oleamide to downregulate gap junction communication in nontransgenic outgrowth cultures showed that this independent method of reducing gap junction communication in cardiac crest cells also resulted in a reduction in the rate of crest migration. To determine the possible relevance of these findings to neural crest migration in vivo, a lacZ transgene was used to visualize the distribution of cardiac neural crest cells in the outflow tract. These studies showed more lacZ-positive cells in the outflow septum in the CMV43 transgenic mice, while a reduction was observed in the α1 connexin KO mice. Surprisingly, this was accompanied by cell proliferation changes, not in the cardiac neural crest cells, but in the myocardium— an elevation in the CMV43 mice vs. a reduction in the α1 connexin KO mice. The latter observation suggests that cardiac neural crest cells may have a role in modulating growth and development of non–neural crest– derived tissues. Overall, these findings suggest that gap junction communication mediated by α1 connexins plays an important role in cardiac neural crest migration. Furthermore, they indicate that cardiac neural crest perturbation is the likely underlying cause for heart defects in mice with the gain or loss of α1 connexin function.