Visual system homeobox-1 （vsxl） is important in retinal progenitor proliferation, differentiation, and function maintenance of bipolar cells in vertebrates. Recent study in Xenopus laevis has shown that vsxl 3＇ unt...Visual system homeobox-1 （vsxl） is important in retinal progenitor proliferation, differentiation, and function maintenance of bipolar cells in vertebrates. Recent study in Xenopus laevis has shown that vsxl 3＇ untranslated region （3＇ UTR） can mediate cell-specific translation of vsxl mRNA in the bipolar cells of the retinal inner nuclear layer （INL）. vsxl is also transcribed at the early developmental stages prior to eye formation and its spatiotemporal expression patterns are conserved in all the examined vertebrates. In order to determine whether the vsxl 3＇ UTR has a role in regulating the spatiotemporal expression of vsxl during early embryogenesis, we constructed a vsxl UTR-controlled green fluorescent protein （GFP） reporter gene system and examined the GFP expression pattern in goldfish, Carassius auratus, at different developmental stages, Our results indicated that both the vsxl 5＇ UTR and the vsxl 3＇ UTR remarkably repressed GFP expression at transcription level but did not regulate tissue-specific translation at early developmental stages. GFP protein was ubiquitously expressed in the embryos injected with GFP-sensors containing vsxl UTRs before 60 h post-fertilization （hpf）. From hatching stage （72 hpf） onwards, however, GFP protein was specifically expressed in the bipolar cells of the retinal 1NL in the vsxl 3＇UTR-GFP-sensor embryos, but was still ubiquitously expressed in the embryos injected with GFP-sensor lacking vsxl 3＇ UTR. These observations showed a significant difference of vsxl 3＇ UTR-mediated translation between early and late developmental stages and suggested that vsxl 3＇ UTR might not be involved in regulating the spatiotemporal expression of vsxl until hatching stage during embryogenesis.展开更多
vsx1(visual system homeobox-1),a homeobox gene originally identified from an adult goldfish retinal cDNA library,has been shown to regulate retina progenitor proliferation,differentiation and functional maintenance of...vsx1(visual system homeobox-1),a homeobox gene originally identified from an adult goldfish retinal cDNA library,has been shown to regulate retina progenitor proliferation,differentiation and functional maintenance of bipolar cells in vertebrates.However,in all the examined vertebrate species,vsx1 transcripts can be also detected at the early developmental stage,suggesting that it may play an important role in regulating early embryogenesis as well.Here,we investigated the function of vsx1 in early embryogenesis of goldfish(Carassius auratus) with both overexpression and gene knockdown approaches.It was found that vsx1 overexpression specifically blocked dorsal midline structure for-mation and vsx1 knockdown led to disorganized dorsal midline structure.Whole-mount in situ hy-bridization revealed that the midline expression of ntl,a key regulatory gene for chordamesoderm,was repressed by vsx1 overexpression but enhanced in the vsx1 knockdown.Furthermore,VSX1 protein could bind ntl promoter directly and was sufficient to inhibit ntl promoter-driven reporter gene green fluorescence protein transcription.Together,these results suggested that vsx1 may act to repress ec-topic expression of ntl in neural progenitor cells to ensure neural tube development in a spatially coordinated pattern during early embryogenesis.展开更多
β-catenin gene is essential for the formation of normal dorsal axial structure in vertebrates. Recent studies have provided evidence that β-catenin has a certain role in restricting the amount of organizer-induced n...β-catenin gene is essential for the formation of normal dorsal axial structure in vertebrates. Recent studies have provided evidence that β-catenin has a certain role in restricting the amount of organizer-induced neurectoderm formation in zebrafish and Xenopus laevis. To further understand how β-catenin represses the induction of neurectoderm formation and whether the inhibition of neural progenitor fates is essential for the normal organizer formation, it was investigated whether β-catenin was involved in repressing the early neural regulatory gene Visual system homeobox-1 (vsx1) expression during embryogenesis. It was observed that functional inhibition of endogenous β-catenin by morpholino oligonucleotides (MO) resulted in extensive and ubiquitous vsx1 expression, β-catenin could repress vsx1 promoter-driven GFP expression and transcriptional factor gene boz, the direct target gene of β-catenin, could directly repress vsx1 transcription by binding to vsx1 proximal promoter. These observations indicate that β-catenin can repress vsx1 ectopic expression cell-autonomously while it activates chordamesodermal genes expression in the precursory cells of chordamesoderm, suggesting both activation of chordamesodermal genes and repression of neural regulatory genes vsx1 in precursory chordamesoderm by β-catenin are essential for normal notochord formation.展开更多
基金supported by the National Science Foundation of China (No.30430370)the State Key Basic Research Project of China (No.2004CB117401)
文摘Visual system homeobox-1 （vsxl） is important in retinal progenitor proliferation, differentiation, and function maintenance of bipolar cells in vertebrates. Recent study in Xenopus laevis has shown that vsxl 3＇ untranslated region （3＇ UTR） can mediate cell-specific translation of vsxl mRNA in the bipolar cells of the retinal inner nuclear layer （INL）. vsxl is also transcribed at the early developmental stages prior to eye formation and its spatiotemporal expression patterns are conserved in all the examined vertebrates. In order to determine whether the vsxl 3＇ UTR has a role in regulating the spatiotemporal expression of vsxl during early embryogenesis, we constructed a vsxl UTR-controlled green fluorescent protein （GFP） reporter gene system and examined the GFP expression pattern in goldfish, Carassius auratus, at different developmental stages, Our results indicated that both the vsxl 5＇ UTR and the vsxl 3＇ UTR remarkably repressed GFP expression at transcription level but did not regulate tissue-specific translation at early developmental stages. GFP protein was ubiquitously expressed in the embryos injected with GFP-sensors containing vsxl UTRs before 60 h post-fertilization （hpf）. From hatching stage （72 hpf） onwards, however, GFP protein was specifically expressed in the bipolar cells of the retinal 1NL in the vsxl 3＇UTR-GFP-sensor embryos, but was still ubiquitously expressed in the embryos injected with GFP-sensor lacking vsxl 3＇ UTR. These observations showed a significant difference of vsxl 3＇ UTR-mediated translation between early and late developmental stages and suggested that vsxl 3＇ UTR might not be involved in regulating the spatiotemporal expression of vsxl until hatching stage during embryogenesis.
基金Supported by National Natural Science Foundation of China (Grant No.30430370)National Key Basic Research and Development Program of China (Grant No.2004CB117401)Doctoral Fund of Ministry of Education of China (Grant No.20050542002)
文摘vsx1(visual system homeobox-1),a homeobox gene originally identified from an adult goldfish retinal cDNA library,has been shown to regulate retina progenitor proliferation,differentiation and functional maintenance of bipolar cells in vertebrates.However,in all the examined vertebrate species,vsx1 transcripts can be also detected at the early developmental stage,suggesting that it may play an important role in regulating early embryogenesis as well.Here,we investigated the function of vsx1 in early embryogenesis of goldfish(Carassius auratus) with both overexpression and gene knockdown approaches.It was found that vsx1 overexpression specifically blocked dorsal midline structure for-mation and vsx1 knockdown led to disorganized dorsal midline structure.Whole-mount in situ hy-bridization revealed that the midline expression of ntl,a key regulatory gene for chordamesoderm,was repressed by vsx1 overexpression but enhanced in the vsx1 knockdown.Furthermore,VSX1 protein could bind ntl promoter directly and was sufficient to inhibit ntl promoter-driven reporter gene green fluorescence protein transcription.Together,these results suggested that vsx1 may act to repress ec-topic expression of ntl in neural progenitor cells to ensure neural tube development in a spatially coordinated pattern during early embryogenesis.
基金supported by the National Natural Science Foundation of China (Grant No. 30430370)National Key Basic Research and Devel-opment Program of China (Grant No. 2004CB117401)
文摘β-catenin gene is essential for the formation of normal dorsal axial structure in vertebrates. Recent studies have provided evidence that β-catenin has a certain role in restricting the amount of organizer-induced neurectoderm formation in zebrafish and Xenopus laevis. To further understand how β-catenin represses the induction of neurectoderm formation and whether the inhibition of neural progenitor fates is essential for the normal organizer formation, it was investigated whether β-catenin was involved in repressing the early neural regulatory gene Visual system homeobox-1 (vsx1) expression during embryogenesis. It was observed that functional inhibition of endogenous β-catenin by morpholino oligonucleotides (MO) resulted in extensive and ubiquitous vsx1 expression, β-catenin could repress vsx1 promoter-driven GFP expression and transcriptional factor gene boz, the direct target gene of β-catenin, could directly repress vsx1 transcription by binding to vsx1 proximal promoter. These observations indicate that β-catenin can repress vsx1 ectopic expression cell-autonomously while it activates chordamesodermal genes expression in the precursory cells of chordamesoderm, suggesting both activation of chordamesodermal genes and repression of neural regulatory genes vsx1 in precursory chordamesoderm by β-catenin are essential for normal notochord formation.
