Cysteine‐ and glycine‐rich protein 1a is involved in spinal cord regeneration in adult zebrafish
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Abstract In contrast to mammals, adult zebrafish have the ability to regrow descending axons and gain locomotor recovery after spinal cord injury (SCI). In zebrafish, a decisive factor for successful spinal cord regeneration is the inherent ability of some neurons to regrow their axons via (re)expressing growth‐associated genes during the regeneration period. The nucleus of the medial longitudinal fascicle (NMLF) is one of the nuclei capable of regenerative response after SCI. Using microarray analysis with laser capture microdissected NMLF, we show that cysteine‐ and glycine‐rich protein (CRP)1a (encoded by the csrp1a gene in zebrafish), the function of which is largely unknown in the nervous system, was upregulated after SCI. In situ hybridization confirmed the upregulation of csrp1a expression in neurons during the axon growth phase after SCI, not only in the NMLF, but also in other nuclei capable of regeneration, such as the intermediate reticular formation and superior reticular formation. The upregulation of csrp1a expression in regenerating nuclei started at 3 days after SCI and continued to 21 days post‐injury, the longest time point studied. In vivo knockdown of CRP1a expression using two different antisense morpholino oligonucleotides impaired axon regeneration and locomotor recovery when compared with a control morpholino, demonstrating that CRP1a upregulation is an important part of the innate regeneration capability in injured neurons of adult zebrafish. This study is the first to demonstrate the requirement of CRP1a for zebrafish spinal cord regeneration.Keywords:
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背景 Tbxl 是为 DiGeorge 症候群(DGS ) 的主要候选人基因。类似于在 DGS 病人，观察的缺点在 Tbxl-/- 动物模型破坏的结构在开发期间从神经的冠房间被导出。尽管一些 Tbxl 击倒的动物模型的词法显型是描述的井，心脏的表演的分析是有限的。因此，心肌的性能在 Tbxl morpholino 被探索注射 zebrafish 胚胎。阐明的方法这些问题， Tbxl 特定的 morpholino 被用来在 zebrafish 减少 Tbxl 的功能。心肌的房间的区别在 situ 杂交用整个山被观察。心率从 24 ～ 72 小时柱子授精(hpf ) 在显微镜下面被观察并且记录。心脏的表演被测量室的弄短的部分和 atrial 弄短 fraction.Results Tbxl morpholino 分析注射胚胎被缺点在咽头的拱门，耳的泡，大动脉的拱门和胸腺描绘。另外， Tbxl 敲击在下面在 zebrafish 减少了咽头的神经的冠房间的数量。反常心脏的形态学在将近 20% Tbxl morpholino 是可见的注射胚胎。在这些胚胎的心没不完全地循环或循环。重要地，心脏的性能和心率在注射 embryos.Conclusions Tbxl 可能玩的 Tbxl morpholino 被减少在在 zebrafish 的咽头的神经的冠房间的开发的一个必要角色。心脏的性能被 Tbxl 损害在 zebrafish 击倒。
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The insulin degrading enzyme (IDE) has a key role in Alzheimer's disease (AD) through its degradation of Amyloid beta peptide (Ab). Interest is growing in using zebrafish as an in vivo model to investigate the function of AD-related genes. The zebrafish IDE sequence shares 89% amino acid residue identity with human IDE and so may possess similar activity. We examined the expression pattern of IDE during zebrafish embryogenesis by in-situ hybridization. We also performed a knockdown study to investigate the IDE function in zebrafish embryos using translation-blocking antisense morpholino. Strong expression of IDE gene in the brain region was observed in zebrafish embryos. Reduction of IDE expression generated phenotypic changes characterized by severe hydrocephalus and cardiac edema and/or a curved trunk. Development of the nervous system appeared to be disturbed as shown by deficits in touch-elicited reflexes. The severe phenotype associated with loss of IDE function in zebrafish indicates that this protein plays an essential role in zebrafish development. Our results provide insights into the function of IDE during zebrafish embryogenesis that have implications for the potential of zebrafish as an AD model.
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Lissencephaly，严重疾病被大脑畸形性描绘。主要原因的基因 lissencephaly 是 LIS1。LIS1 的变化或删除在大脑开发导致增长和神经原的迁居缺乏。然而，很少在胚胎的开发对它的生物函数被知道。在这篇文章，我们识别了 zebrafish LIS1 基因的表达式模式并且在胚胎的开发调查了它的函数。我们证明 zebrafish 由二 LIS1 基因， LIS1a 和 LIS1b 组成了。生物信息学分析表明 LIS1 基因在蛋白质序列和 genomic 结构两个都在进化被保存。zebrafish LIS1a 和 LIS1b 的表示模式证明两个抄本无所不在地被表示根本胚胎的发展阶段并且在检验的成年纸巾。在蛋白质水平， LIS1 产品主要在由西方的弄污显示出分析的早阶段在大脑织物并且在胚胎存在。整个山的 immunostaining 数据证明 LIS1 蛋白质在整个从 1 房间舞台的胚胎是分布式的到 5 白天授精以后。通过 morpholino antisense oligonucleotides 的 LIS1 蛋白质表示击倒在 zebrafish，包括的大脑畸形性，发行量反常，和身体导致了许多发展缺乏卷屈。一起拿，我们的学习建议 zebrafish LIS1 在胚胎的开发起一个很重要的作用。
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MicroRNAs are an important class of gene expression regulators that are involved in many biological processes including embryogenesis. miR-125b is a conserved microRNA that is enriched in the nervous system. We have previously reported the function of miR-125b in neuronal differentiation of human cell lines. We also discovered the function of miR-125b in regulating p53 in human and zebrafish. Here we further characterize the brain defects in zebrafish embryos injected with morpholinos against miR-125b. Our data confirm the essential role of miR-125b in brain morphogenesis particularly in maintaining the balance between proliferation, cell death and differentiation. We identified lunatic fringe (lfng) as an additional target of miR-125b in human and zebrafish and suggest that lfng may mediate the function of miR-125b in neurogenesis. Together, this report reveals new insights into the function of miR125b during neural development of zebrafish. Keywords—microRNA, miR-125b, neurogenesis, zebrafish.
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Dihydropyrimidinase-like family proteins (Dpysls) are relevant in several processes during nervous system development; among others, they are involved in axonal growth and cell migration. Dpysl2 (CRMP2) is the most studied member of this family; however, its role in vivo is still being investigated. Our previous studies in zebrafish showed the requirement of Dpysl2 for the proper positioning of caudal primary motor neurons and Rohon-Beard neurons in the spinal cord.In the present study, we show that Dpysl2 is necessary for the proper migration of facial branchiomotor neurons during early development in zebrafish. We generated a dpysl2 knock-out (KO) zebrafish mutant line and used different types of antisense morpholino oligonucleotides (AMO) to analyze the role of Dpysl2 in this process. Both dpysl2 KO mutants and morphants exhibited abnormalities in the migration of these neurons from rhombomers (r) 4 and 5 to 6 and 7. The facial branchiomotor neurons that were expected to be at r6 were still located at r4 and r5 hours after the migration process should have been completed. In addition, mutant phenotypes were rescued by injecting dpysl2 mRNA into the KO embryos. These results indicate that Dpysl2 is involved in the proper migration of facial branchiomotor neurons in developing zebrafish embryos.
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MicroRNAs are an important class of gene expression regulators that are involved in many biological processes including embryogenesis. miR-125b is a conserved microRNA that is enriched in the nervous system. We have previously reported the function of miR-125b in neuronal differentiation of human cell lines. We also discovered the function of miR-125b in regulating p53 in human and zebrafish. Here we further characterize the brain defects in zebrafish embryos injected with morpholinos against miR-125b. Our data confirm the essential role of miR-125b in brain morphogenesis particularly in maintaining the balance between proliferation, cell death and differentiation. We identified lunatic fringe (lfng) as an additional target of miR-125b in human and zebrafish and suggest that lfng may mediate the function of miR-125b in neurogenesis. Together, this report reveals new insights into the function of miR125b during neural development of zebrafish. Keywords—microRNA, miR-125b, neurogenesis, zebrafish.
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