乙醇对斑马鱼胚胎发育中tenascin-c基因表达的影响

Significance of Expression of Tenascin-c Gene in Zebrafish Embryo Development Induced by Ethanol

目的探讨tenascin-c(tnc)基因在斑马鱼胚胎异常发育过程中表达的变化,从而揭示该基因在斑马鱼胚胎发育中可能存在的作用。方法不同水平乙醇(100、200、300、400、500mmol/L)持续处理斑马鱼胚胎,制备斑马鱼异常发育模型,收集受精后24、48h斑马鱼胚胎并固定,用原位杂交和反转录聚合酶链反应(RT-PCR)方法观察tnc在斑马鱼异常发育时表达的变化。组间比较采用SPSS11.5软件进行统计学处理。结果200mmol/L乙醇处理的斑马鱼胚胎在受精72h其他系统未发现发育异常,而心脏已出现畸形。100、200mmol/L乙醇处理后,RT-PCR结果示tnc在斑马鱼胚胎发育中的表达上调,而原位杂交信号变化不明显;300mmol/L及以上乙醇处理的胚胎,原位杂交结果示tnc在体节处表达下降;100～400mmol/L乙醇处理的胚胎,在受精48htnc在斑马鱼心脏部位均有表达。结论tnc在低水平乙醇处理时表达即上调,对胚胎正常发育有保护作用;在心脏异常发育时表达,促进心脏的正常发育;tnc在病理状态下的表达模式不仅在脊椎动物中具有保守性,而且在胚胎和成体时期也具有保守性。

Objective To explore the expression pattern of tenascin - c（mc） gene in zebrafish embryo abnormal development which was induced by ethanol,and to further understand the function of tnc gene in embryo develepment. Methods Zebrafish were treated with ethanol at different concentration from 100 to 500 mmol/L,and embryos at 24 and 48 hours were collected and fixed,then tnc expression pattern was observed by in situ hybridization and reverse transcriptase - polymerase chain reaction（ RT - PCR）. Results The result of RT - PCR showed that ethanol at 100 and 200 mmol/L could increase the expression of tnc, while the result of in situ hybridization showed that,while ethanol at 300 mmol/L and above decrease the expression of mc in presumptive position at 24 hours, and ethanol at 100 mmol/L and above caused increase expression of mc in zebrafish heart. Conclusions mc is increased when treated with 100 and 200 mmol/L ethanol and is presented in the abnormal development of hearts of zebrafish,which can promote the normal development of embryos in some degrees. The expression pattern of tnc in pathologic state is highly conserved in all vertebrate,and in adult and embryos as well.