OAZ在果蝇大脑发育中的功能研究
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摘要
OAZ基因编码在进化上保守的C2H2型O/E相关锌指蛋白,参与基因转录的调控。TGF-β信号传导通路中OAZ通过与SMAD4/R-SMAD结合形成转录复合物发挥作用;Olf1/EBF作为转录因子与OAZ相互作用来调控嗅觉上皮细胞和B淋巴细胞的分化。此外,OAZ是JAK/STAT信号通路的下游候选靶基因。但是迄今为止OAZ在果蝇大脑发育的功能还没有研究。果蝇大脑视叶作为一个相对简易操作的模型为我们揭示早期神经干细胞的增殖和转化机制创造了条件,为加快理解哺乳动物早期神经发生过程以及进一步开展神经干细胞治疗提供可能。本研究通过RNA干扰来研究OAZ在果蝇大脑发育中的作用。我们的初步实验结果表明,OAZ对果蝇大脑视叶神经上皮细胞的维持可能不是必需的,OAZ对果蝇大脑视叶神经板和脑髓神经节的发育也可能不是必需的。
The OAZ gene encoded an evolutionarily conserved C2 H2 O/E-related zinc finger protein, which was involved in the regulation of gene transcription. In the TGF-β signaling pathway, OAZ interacted with SMAD4/R-SMAD to form transcriptional complexes; Olf1/EBF acted as a transcription factor to interact with OAZ to regulate the differentiation of olfactory epithelial cells and B lymphocytes. In addition, OAZ was a candidate target gene of the JAK/STAT signaling pathway. However, the role of OAZ in Drosophila brain development has not been investigated. The Drosophila larval optic lobe as a model, which was relatively simple to operate created a condition for us to reveal the multiplication and transfer mechanism of early neural stem cells. And it was possible to accelerate the understanding of early neurogenesis in mammals and to further develop neural stem cell therapy. Here the role of the OAZ gene in the regulation of neural stem cells was examined through knockdown of OAZ m RNA. Our data suggested that OAZ might not be necessary to maintain the neuroepithelia in Drosophila optic lobes, and OAZ might not be necessary for the development of lamina and medulla, two visual processing centers in the brain.
The OAZ gene encoded an evolutionarily conserved C2 H2 O/E-related zinc finger protein, which was involved in the regulation of gene transcription. In the TGF-β signaling pathway, OAZ interacted with SMAD4/R-SMAD to form transcriptional complexes; Olf1/EBF acted as a transcription factor to interact with OAZ to regulate the differentiation of olfactory epithelial cells and B lymphocytes. In addition, OAZ was a candidate target gene of the JAK/STAT signaling pathway. However, the role of OAZ in Drosophila brain development has not been investigated. The Drosophila larval optic lobe as a model, which was relatively simple to operate created a condition for us to reveal the multiplication and transfer mechanism of early neural stem cells. And it was possible to accelerate the understanding of early neurogenesis in mammals and to further develop neural stem cell therapy. Here the role of the OAZ gene in the regulation of neural stem cells was examined through knockdown of OAZ m RNA. Our data suggested that OAZ might not be necessary to maintain the neuroepithelia in Drosophila optic lobes, and OAZ might not be necessary for the development of lamina and medulla, two visual processing centers in the brain.
引文
Betschinger J.,Mechtler K.,and Knoblich J.A.,2006,Asymmetric segregation of the tumor suppressor brat regulates self renewal in Drosophila neural stem cells,Cell,124(6):1241-1253
Brand A.H.,and Livesey F.J.,2011,Neural stem cell biology in vertebrates and invertebrates:more alike than different,Neuron,70(4):719-729
Egger B.,Boone J.Q.,Steveens N.R.,Brand A.H.,and Doe C.Q.,2007,Regulation of spindle orientation and neural stem cell fate in the Drosophila optic lobe,Neural Development,2:1
Eroglu E.,Burkard T.R.,Jiang R.,Saini N.,Homem C.C.F.,Reichert H.,and Knoblich J.A.,2014,SWI/SNF complex prevent lineage reversion and induces temporal patterning in neural stem cells,Cell,156(6):1259-1273
Farkas L.M.,and Huttner W.B.,2008,The cell biology of neural stem and progenitor cells and its significance for their proliferation versus differentiation during mammalian brain development,Current Opinion in Cell Biology,20(6):707-715
Feng X.B.,Li R.L.,Huang J.,Zhang H.Y.,Zhu L.N.,Hua B.Z.,Tsao B.P.,and Sun L.Y.,2010,Olf1/EBF associated zinc finger protein interfered with antinuclear antibody production in patients with systemic lupus erythematosus,Arthritis Research and Therapy,12(2):R59
Feng X.B.,Shen N.,Chen S.L.,Qian J.,Wu H.,Bao C.D.,and Wang Y.,2003,Susceptibility gene of systemic lupus erythematosus in 16q12 in Chinese cohort,Zhongguo Yixue Yichuan Zazhi(Chinese Journal of Medical Genetics),20(1):27-30(冯学兵,沈南,陈顺乐,钱捷,吴慧,鲍春德,王元,2003,中国人群16号染色体q12区存在系统性红斑狼疮易感基因,中国医学遗传杂志,20(1):27-30)
Feng Y.,and Shao Z.T.,2012,Investigation of roles of ion channel proteins in the development of neuroepithelial stem cells,Jiguang Shengwu Xuebao(Acta Laser Biology Sini ca),21(4):310-315(冯艳,邵占涛,2012,离子通道蛋白在果蝇大脑神经上皮干细胞发育中的功能研究,激光生物学报,21(4):310-315)
Got M.,and Huttner W.B.,2007,The cell biology of neurogenesis,Nature Reviews Molecular Cell Biology,6(10):777-788
Harrison M.M.,Li X.Y.,Kaplan T.,Botchan M.R.,and Eisen M.B.,2011,Zelda binding in the early Drosophila mela-%nogaster embryo marks regions subsequently activated at the maternal-to-zygotic transition,PLo S Genetics,7(10):e1002266
Hata A.,Seoane J.,Lagna G.,Montalvo E.,Hemmati-Brivanlou A.,and Massague J.,2000,OAZ uses distinct DNA and protein binding zinc fingers in separate BMP-Smad and Olf signaling pathways,Cell,100(2):229-240
Hrdlicka L.,Gibson M.,Kiger A.,Micchelli C.,Schober M.,Schock F.,and Perrimon N.,2002,Analysis of twentyfour Gal4 lines in Drosophila melanogaster,Genesis,34(1-2):51-57
Hunter C.M.,Huang W.,Mackay T.F.,and Singh N.D.,2016,The genetic architecture of natural variation in recombination rate in Drosophila melanogaster,Public Library of Science Genetics,12(4):e1005951
Ku M.,Howard S.,Ni W.,Lagna G.,and Hata A.,2006,OAZregulates bone morphogenetic protein signaling through Smad6 activation,Journal of Biological Chemistry,281(8):5277-5287
Lee H.S.,Lee S.Y.,Lee H.Hwang Y.S.,Cha S.W.,Park S.,Lee J.Y.,Park J.B.,Kim S.,Park M.J.,and Kim J.,2011,Direct response elements of BMP within the PV.1A promoter are essential for its transcriptional regulation during early Xenopus development,Public Library of Science One,6(8):e22621
Liang D.,Huang X.Q.,Shen N.,Mao H.Q.,Feng X.B.,Huang X.F.,Tang J.P.,Chen X.X.,Chen S.L.,Gu Y.Y.,Bao C.D.,Wang Y.,and Qian J.,2005,Variations within OLF1_EBF-associated zinc finger protein gene confer susceptibility to lupus nephritis in Chinese population,Zhongguo Yixue Zazhi(National Medical Journal of China),85(14):949-954(梁栋,黄秀琴,沈南,毛寒倩,冯学兵,黄新芳,汤建平,陈晓翔,陈顺乐,顾越英,鲍春德,王元,钱捷,2005,早期B细胞因子相关锌指蛋白基因的变异与中国人群的狼疮肾炎发病的相关性研究,中国医学杂志,85(14):949-954)
Manseau L.,Baradaran A.,Brower D.,Budhu A.,Elefant F.,Plan H.,Philip A.V.,Yang M.,Glover D.,Kaiser K.,Palter K.,and Selleck S.,1997,Gal4 enhancer traps expressed in the embryo,larval brain,imaginal discs,and ovary of Drosophi-%la,Developmental Dynamics,209(3):310-322
Morrison S.J.,and Kimble J.,2006,Asymmetric and symmetric stem-cell divisions in development and cancer,Nature,441(7097):1068-1074
Ngo K.T.,Wang J.,Junker M.,Kri Z S.,Vo G.,Asem B.,Olson J.M.,Banerjee U.,and Hartenstein V.,2010,Concomitant requirement for Notch and JAK/STAT signaling during neuroepithelial differentiation in the Drosophila optic lobe1,Biochemical Biophysical Research Communications,346(2):284-295
Schulz K.N.,Bondra E.R.,Moshe A.,Villalta J.E.,Lieb J.D.,Kaplan T.,McKay D.J.,and Harrison M.M.,2015,Zelda is differentially required for chromatin accessibility,transcription factor binding,and gene expression in the early Drosophila embryo,Genome Research,25(11):1715-1726
Temple S.,2001,The development of neural stem cells,Nature,414(6859):112-117
Wang H.B.,Chen X.,He T.,Zhou Y.N.,and Luo H.,2013,Evidence for tissue-specific JAK/STAT target genes in Drosophila optic lobe development,Genetics,195(4):1291-1306
Wang W.,Liu W.,Wang Y.,Zhou L.,Tang X.,and Luo H.,2011,Notch signaling regulates neuroepithelial stem cell maintenance and neuroblast formation in Drosophila optic lobe development,Developmental Biology,350(2):414-428
Weissman I.L.,Anderson D.J.,and Gage F.,2001,Stem and progenitor cell:Origins,phenotypes,lineage commitments,and trans differentiations,Annual Review of Cell and Developmental Biology,17:387-403
Wrana J.L.,2000,Regulation of smad activity,Cell,100(2):189-192
Brand A.H.,and Livesey F.J.,2011,Neural stem cell biology in vertebrates and invertebrates:more alike than different,Neuron,70(4):719-729
Egger B.,Boone J.Q.,Steveens N.R.,Brand A.H.,and Doe C.Q.,2007,Regulation of spindle orientation and neural stem cell fate in the Drosophila optic lobe,Neural Development,2:1
Eroglu E.,Burkard T.R.,Jiang R.,Saini N.,Homem C.C.F.,Reichert H.,and Knoblich J.A.,2014,SWI/SNF complex prevent lineage reversion and induces temporal patterning in neural stem cells,Cell,156(6):1259-1273
Farkas L.M.,and Huttner W.B.,2008,The cell biology of neural stem and progenitor cells and its significance for their proliferation versus differentiation during mammalian brain development,Current Opinion in Cell Biology,20(6):707-715
Feng X.B.,Li R.L.,Huang J.,Zhang H.Y.,Zhu L.N.,Hua B.Z.,Tsao B.P.,and Sun L.Y.,2010,Olf1/EBF associated zinc finger protein interfered with antinuclear antibody production in patients with systemic lupus erythematosus,Arthritis Research and Therapy,12(2):R59
Feng X.B.,Shen N.,Chen S.L.,Qian J.,Wu H.,Bao C.D.,and Wang Y.,2003,Susceptibility gene of systemic lupus erythematosus in 16q12 in Chinese cohort,Zhongguo Yixue Yichuan Zazhi(Chinese Journal of Medical Genetics),20(1):27-30(冯学兵,沈南,陈顺乐,钱捷,吴慧,鲍春德,王元,2003,中国人群16号染色体q12区存在系统性红斑狼疮易感基因,中国医学遗传杂志,20(1):27-30)
Feng Y.,and Shao Z.T.,2012,Investigation of roles of ion channel proteins in the development of neuroepithelial stem cells,Jiguang Shengwu Xuebao(Acta Laser Biology Sini ca),21(4):310-315(冯艳,邵占涛,2012,离子通道蛋白在果蝇大脑神经上皮干细胞发育中的功能研究,激光生物学报,21(4):310-315)
Got M.,and Huttner W.B.,2007,The cell biology of neurogenesis,Nature Reviews Molecular Cell Biology,6(10):777-788
Harrison M.M.,Li X.Y.,Kaplan T.,Botchan M.R.,and Eisen M.B.,2011,Zelda binding in the early Drosophila mela-%nogaster embryo marks regions subsequently activated at the maternal-to-zygotic transition,PLo S Genetics,7(10):e1002266
Hata A.,Seoane J.,Lagna G.,Montalvo E.,Hemmati-Brivanlou A.,and Massague J.,2000,OAZ uses distinct DNA and protein binding zinc fingers in separate BMP-Smad and Olf signaling pathways,Cell,100(2):229-240
Hrdlicka L.,Gibson M.,Kiger A.,Micchelli C.,Schober M.,Schock F.,and Perrimon N.,2002,Analysis of twentyfour Gal4 lines in Drosophila melanogaster,Genesis,34(1-2):51-57
Hunter C.M.,Huang W.,Mackay T.F.,and Singh N.D.,2016,The genetic architecture of natural variation in recombination rate in Drosophila melanogaster,Public Library of Science Genetics,12(4):e1005951
Ku M.,Howard S.,Ni W.,Lagna G.,and Hata A.,2006,OAZregulates bone morphogenetic protein signaling through Smad6 activation,Journal of Biological Chemistry,281(8):5277-5287
Lee H.S.,Lee S.Y.,Lee H.Hwang Y.S.,Cha S.W.,Park S.,Lee J.Y.,Park J.B.,Kim S.,Park M.J.,and Kim J.,2011,Direct response elements of BMP within the PV.1A promoter are essential for its transcriptional regulation during early Xenopus development,Public Library of Science One,6(8):e22621
Liang D.,Huang X.Q.,Shen N.,Mao H.Q.,Feng X.B.,Huang X.F.,Tang J.P.,Chen X.X.,Chen S.L.,Gu Y.Y.,Bao C.D.,Wang Y.,and Qian J.,2005,Variations within OLF1_EBF-associated zinc finger protein gene confer susceptibility to lupus nephritis in Chinese population,Zhongguo Yixue Zazhi(National Medical Journal of China),85(14):949-954(梁栋,黄秀琴,沈南,毛寒倩,冯学兵,黄新芳,汤建平,陈晓翔,陈顺乐,顾越英,鲍春德,王元,钱捷,2005,早期B细胞因子相关锌指蛋白基因的变异与中国人群的狼疮肾炎发病的相关性研究,中国医学杂志,85(14):949-954)
Manseau L.,Baradaran A.,Brower D.,Budhu A.,Elefant F.,Plan H.,Philip A.V.,Yang M.,Glover D.,Kaiser K.,Palter K.,and Selleck S.,1997,Gal4 enhancer traps expressed in the embryo,larval brain,imaginal discs,and ovary of Drosophi-%la,Developmental Dynamics,209(3):310-322
Morrison S.J.,and Kimble J.,2006,Asymmetric and symmetric stem-cell divisions in development and cancer,Nature,441(7097):1068-1074
Ngo K.T.,Wang J.,Junker M.,Kri Z S.,Vo G.,Asem B.,Olson J.M.,Banerjee U.,and Hartenstein V.,2010,Concomitant requirement for Notch and JAK/STAT signaling during neuroepithelial differentiation in the Drosophila optic lobe1,Biochemical Biophysical Research Communications,346(2):284-295
Schulz K.N.,Bondra E.R.,Moshe A.,Villalta J.E.,Lieb J.D.,Kaplan T.,McKay D.J.,and Harrison M.M.,2015,Zelda is differentially required for chromatin accessibility,transcription factor binding,and gene expression in the early Drosophila embryo,Genome Research,25(11):1715-1726
Temple S.,2001,The development of neural stem cells,Nature,414(6859):112-117
Wang H.B.,Chen X.,He T.,Zhou Y.N.,and Luo H.,2013,Evidence for tissue-specific JAK/STAT target genes in Drosophila optic lobe development,Genetics,195(4):1291-1306
Wang W.,Liu W.,Wang Y.,Zhou L.,Tang X.,and Luo H.,2011,Notch signaling regulates neuroepithelial stem cell maintenance and neuroblast formation in Drosophila optic lobe development,Developmental Biology,350(2):414-428
Weissman I.L.,Anderson D.J.,and Gage F.,2001,Stem and progenitor cell:Origins,phenotypes,lineage commitments,and trans differentiations,Annual Review of Cell and Developmental Biology,17:387-403
Wrana J.L.,2000,Regulation of smad activity,Cell,100(2):189-192