Zhu Xianmin, Liu Rui, Guan Jun, Zeng Wen, Yin Jin, Zhang Yicheng
Department of Medical Oncology, Hubei Cancer Hospital, Wuhan, Hubei 430079, China.
Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Int J Med Sci. 2017 Jul 18;14(8):758-763. doi: 10.7150/ijms.18907. eCollection 2017.
Zebrafish () are an attractive vertebrate model for the molecular dissection of disease mechanisms. Janus kinase (JAK)/signal transducer and activator of transcription (stat) has been defined through studies of cytokine signaling pathways in mammals. Here, we examined the expression level of Jak2a, which is a homolog of mammalian jak2 in zebrafish, by quantitative reverse transcriptase (RT)-PCR, and the peak of mRNA expression occurred at 3.75 hours post fertilization (hpf). The overexpression of Jak2a was proven by real-time Q-PCR and Western blot in 1-4-cell stage embryos injected with 400 ng/µl full-length jak2a mRNA as well as gfi1.1, gata1, mpo and β-embryonic hemoglobin as detected by real-time Q-PCR. Moreover, jak2a mRNA significantly increased the GFP population in the transgenic zebrafish lines Tg () (uninjected embryos: 17.22±1.70%; embryos injected with jak2a mRNA: 21.31±2.11%, p<0.01) and Tg () (uninjected embryos: 3.86±1.94; embryos injected with jak2a mRNA: 6.64±1.30%, p<0.01) compared with the control group. Thus, our data indicate that Jak2a plays an important role in erythropoiesis and myeloid hematopoiesis.
斑马鱼()是用于分子剖析疾病机制的一种有吸引力的脊椎动物模型。通过对哺乳动物细胞因子信号通路的研究,已明确了Janus激酶(JAK)/信号转导子和转录激活子(STAT)。在此,我们通过定量逆转录酶(RT)-PCR检测了斑马鱼中哺乳动物jak2的同源物Jak2a的表达水平,mRNA表达峰值出现在受精后3.75小时(hpf)。通过实时定量PCR和蛋白质免疫印迹法证实,在注射400 ng/µl全长jak2a mRNA以及gfi1.1、gata1、mpo和β-胚胎血红蛋白的1-4细胞期胚胎中,Jak2a过表达,实时定量PCR检测到了这些基因。此外,与对照组相比,jak2a mRNA显著增加了转基因斑马鱼品系Tg()(未注射胚胎:17.22±1.70%;注射jak2a mRNA的胚胎:21.31±2.11%,p<0.01)和Tg()(未注射胚胎:3.86±1.94;注射jak2a mRNA的胚胎:6.64±1.30%,p<0.01)中的绿色荧光蛋白群体。因此,我们的数据表明Jak2a在红细胞生成和髓系造血中起重要作用。