[Myeloid and erythroid hematopoietic transcription factor expression decline after knockdown of lmna genes in zebrafish embryos].
[Myeloid and erythroid hematopoietic transcription factor expression decline after knockdown of lmna genes in zebrafish embryos].
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斑马鱼胚胎中lmna基因敲低后骨髓和红系造血转录因子表达下降
DOI：
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发表时间：
2016-05
期刊：
影响因子：
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通讯作者：
Shu Liping
中科院分区：
文献类型：
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作者：
Hou Shifang;Wang Zhihua;Wang Jun;He Zhixu;Shu Liping
目的：探讨lmna基因下调对斑马鱼早期造血发育的影响。方法：采用磷酸二酰胺吗啉低聚物（PMO）技术下调斑马鱼lmna基因表达。以单细胞阶段显微注射lmna基因mRNA的磷酸二酰胺吗啉反义寡核苷酸的斑马鱼胚胎为实验组，注射无意义的磷酸二酰胺吗啉反义寡核苷酸的斑马鱼胚胎作为对照。受精后18、24、30和36 hpf收集胚胎。采用实时荧光定量PCR（RT-PCR）和全胚原位杂交方法检测髓系造血转录因子pu的表达。 1 和斑马鱼红系造血转录因子 gata1。结果：RT-PCR显示pu．与对照组相比，实验组gata1和1均下降（均P<0.05）。全胚原位杂交显示pu的蓝黑阳性杂交信号。实验组的gata1和1均较对照组浅。结论：斑马鱼的髓系造血和红系造血由于lmna基因的下调而受阻。
Objective: To investigate the effect of lmna gene down-regulation on early hematopoietic development of zebrafish. Methods: Phosphorodiamidate morpholino oligomer (PMO) technology was used to downregulate lmna gene expression in Zebrafish. Zebrafish embryos injected phosphorodiamidate morpholino antisense oligonucleotide of lmna gene mRNA by microinjection at unicellular stage were taken as the experimental group, and those injected meaningless phosphorodiamidate morpholino antisense oligonucleotide were taken as the control. The embryos were collected at 18, 24, 30 and 36 hpf after the fertilization. The real-time fluorescent quantitative PCR (RT-PCR) and whole embryo in situ hybridization methods were used to detect the expression of myeloid hematopoietic transcription factor pu. 1 and erythroid hematopoietic transcription factor gata1 in zebrafish. Results: RT-PCR showed that the expressions of pu. 1 and gata1 decreased in the experimental group compared with the control group (all P<0.05). Whole embryo in situ hybridization showed that the blue-black positive hybridization signals of pu. 1 and gata1 in experimental group were shallow than those in the control group. Conclusion: Myeloid hematopoietic and erythroid hematopoietic of zebrafish are blocked with the downregulation of lmna gene.