miR-30c过表达对斑马鱼早期心脏发育的影响  

Effects of miR - 30c over - expression on early cardiac development of zebrafish

作　　者：徐晶[1] 李萌萌[1] 刘雪华[1] 余章斌[1] 朱春[1] 韩树萍[1] 胡晓山[1] 朱金改[1] 

机构地区：[1]南京医科大学附属南京妇幼保健院儿科,210004

出　　处：《中华实用儿科临床杂志》2016年第1期33-36,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81370200）;江苏省自然科学基金（BK20131075）;江苏省医学重点人才基金（RC2011021）

摘　　要：目的探究miR-30c过表达对斑马鱼早期心脏发育的影响。方法于斑马鱼胚胎1细胞受精卵期，显微注射3个不同浓度梯度的miR-30c模拟物以实现过表达。通过计数在不同时间点斑马鱼胚胎的病死率及心率、显微镜观察斑马鱼大体形态及心包水肿情况，以及胚胎整体原位杂交等方法，评价miR-30c过表达对斑马鱼早期心脏发育的影响。结果与野生型对照组比较，随着过表达浓度的提高，斑马鱼胚胎的病死率逐渐增高，至8μmol／L浓度时，斑马鱼受精后96h病死率已达80％。而同时，作为评价心脏功能重要指标的心率逐渐下降。此外，随着过表达浓度的递增，斑马鱼心包水肿的程度越来越高，且此种心包水肿随着时间的推移越来越严重。之后，通过胚胎整体原位杂交的方法，观察心脏部位特异性基因的表达，发现过表达miR-30c的斑马鱼中，标记心房肌及心室肌探针的表达范围都有所扩大，心脏环化受到抑制，且心脏房室通道瓣膜处的基因表达缺失。结论miR-30c过表达影响斑马鱼的早期心脏发育。Objective To explore the effects of miR - 30c over - expression on early cardiac development of zebrafish. Methods The single - cell - stage zebrafish embryos were micro - injected with 3 different concentrations of miR - 30c mimics in order to model the miR - 30c over - expressed animal. Besides, the effects of miR - 30c overex- pression on cardiac development were investigated. The mortality rate and the heart rate were assessed. Beside, the gross morphology and heart morphology of the zebrafish were observed. Moreover, the molecular mechanisms underlying miR -30c function in zebrafish cardiac development was explored ,and in -situ hybridization was performed. Results Compared with the wild - type embryos, the mortality rate of zebrafish was increased with the rising miR - 30c concen- tration. Furthermore,when the miR- 30c mimic concentration was up to 8 txmol/L, the mortality rate reached 80% at 96 h post - fertilization. Simultaneously, the heart rate, which was viewed as an important index to cardiac function, was decreased. Moreover, the pericardial edema was getting worse over time with increasing concentration of overexpression. Then,the cardiac specific gene expression on the zebrafish embryos by whole - mount in situ hybridization was ex- plored. The area of the cardiac chamber was extended and the heart tube looping was negatively affected. Besides, the expression of atrioventricular canal marker genes was absent in zebrafish embryos from miR - 30c over - expression groups. Conclusion miR -30c can impact the early cardiac development of zebrafish.

关 键 词：miR-30c 斑马鱼 心脏发育 

分 类 号：R714.5[医药卫生—妇产科学]