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Developmental expression of the N-myc downstream regulated gene (Ndrg) family during Xenopus tropicalis embryogenesis. 热带非洲爪蟾胚胎发生过程中N-myc下游调控基因(Ndrg)家族的发育表达。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.150178xh
Chao Zhong, Yan-Kuan Zhou, Shan-shan Yang, Jun-Fang Zhao, Xiao-long Zhu, Hen-Huang Chen, Peichao Chen, Liquan Huang, Xiao Huang
The N-myc downstream regulated gene (Ndrg) family consists of four main members Ndrg1, 2, 3, and 4. The Ndrg genes are involved in many vital biological events including development. However, comprehensive expression patterns of this gene family during vertebrate embryogenesis remain largely unknown. Here, we analyzed the Ndrg family from the evolutionary perspective and examined the expression patterns of the Ndrg genes during Xenopus tropicalis embryogenesis. Different Ndrg family members of vertebrates are separated into different homology clusters which can be further classified into two groups and each Ndrg family member is well conserved during evolution. The temporal and spatial expression patterns of Ndrg1, 2, 3 and 4 are different during early Xenopus tropicalis development. Ndrg1, 2 and 4 are maternally expressed genes while Ndrg3 is a zygotically expressed gene. The Ndrg genes are differentially expressed in the developing central nervous system, the developing sensory organs, and the developing excretory organs. Moreover, they also show other specific expression domains. Our results indicate that the Ndrg genes exhibit specific expression patterns and may play different roles during vertebrate embryogenesis.
N-myc下游调节基因(Ndrg)家族由四个主要成员Ndrg1、2、3和4组成。Ndrg基因参与了许多重要的生物事件,包括发育。然而,该基因家族在脊椎动物胚胎发生过程中的全面表达模式在很大程度上仍然未知。本文从进化角度分析了热带非洲爪蟾Ndrg家族,并研究了Ndrg基因在热带非洲爪蟾胚胎发生过程中的表达模式。脊椎动物不同的Ndrg家族成员被划分为不同的同源群,可进一步分为两类,每个Ndrg家族成员在进化过程中都有很好的保守性。ndr1、2、3和4在热带爪蟾发育早期的时空表达模式存在差异。Ndrg1、2和4是母系表达基因,而Ndrg3是合子表达基因。Ndrg基因在发育中的中枢神经系统、感觉器官和排泄器官中有差异表达。此外,它们还显示出其他特定的表达域。我们的结果表明,Ndrg基因在脊椎动物胚胎发生过程中表现出特定的表达模式,并可能发挥不同的作用。
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引用次数: 11
T-cell internal antigen 1 counteracts somatic RNA degradation during early Xenopus embryogenesis. 在非洲爪蟾胚胎发生早期,t细胞内抗原1抵消体细胞RNA降解。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.150137db
Diana Bauermeister, Maike Claussen, T. Pieler
In Xenopus laevis, maternal transcripts that localize to the vegetal cortex of the oocyte are specifically inherited by prospective germ cells during cleavage stages. While a large fraction of maternal transcripts is degraded during the maternal to zygotic transition (MZT), transcripts associated with the germ-line are stable. A sequence in the dead end 1 3'UTR mediates vegetal localization in the oocyte as well as miR mediated clearance in somatic cells and germ cell specific stabilization during the MZT in embryos. We could identify Tia1 to co-precipitate with known components of vegetal localization RNPs in X. laevis oocytes. Tia1 interacts and co-localizes with various localization elements from vegetally localizing RNAs. In X. laevis embryos, ectopic expression of Tia1 counteracts somatic degradation of dnd1 localization element reporter RNAs and it can synergize with Dnd1 protein in reporter RNA stabilization. Ectopic Tia1 also protects several endogenous localizing and germ cell specific mRNAs from somatic degradation. Thus, proteins that protect germ-line transcripts from miR mediated decay during the MZT in embryos might bind these RNAs already in the oocyte.
在非洲爪蟾(Xenopus laevis)中,定位于卵母细胞植物皮质的母体转录本在卵裂阶段被未来的生殖细胞特异性地遗传。在母系向合子转化(MZT)过程中,大部分母系转录本被降解,而与种系相关的转录本是稳定的。在胚胎MZT过程中,死端13 ' utr中的一个序列介导卵母细胞的植物定位,以及miR介导的体细胞清除和生殖细胞特异性稳定。我们可以确定Tia1与已知的植物定位RNPs成分在褐家鼠卵母细胞中共沉淀。Tia1与来自植物定位rna的各种定位元件相互作用并共定位。在青竹胚胎中,Tia1的异位表达抵消了dnd1定位元件报告RNA的体细胞降解,并与dnd1蛋白协同稳定报告RNA。异位Tia1还保护几种内源性定位和生殖细胞特异性mrna免受体细胞降解。因此,在胚胎MZT期间保护种系转录本免受miR介导的衰变的蛋白质可能结合这些已经在卵母细胞中的rna。
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引用次数: 0
External ear microRNA expression profiles during mouse development. 小鼠发育过程中外耳microRNA表达谱。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.150124sf
Leda Torres, Ulises Juárez, Laura García, J. Miranda-Ríos, S. Frías
MicroRNAs (miRNAs) comprise a class of approximately 22 nucleotide regulatory non-coding RNAs that play several roles in diverse biological processes. Recent reports suggest that embryonic development in mammals is accompanied by dynamic changes in miRNA expression; however, there is no information regarding the role of miRNAs in the development of the external ear. The aim of this study was to determine the stage-specific expression of miRNAs during mouse external ear development in order to identify potentially implicated miRNAs along with their possible targets. miRNA expression profiles from fetal mice pinnae and back skin tissues at 13.5 dpc and 14.5 dpc were obtained using an Affymetrix GeneChip miRNA 3.0 array. Biological triplicates for both tissues, each collected from a litter averaging 16 fetuses, were analyzed. The results were analyzed with Affymetrix's Transcriptome Analysis Console software to identify differentially expressed miRNAs. We observed differential expression of 40 miRNAs including some predicted to target genes implicated in external ear development, such as mmu-miR-10a, an miRNA known to modulate Hoxa1 mRNA levels, and mmu-miR-200c and mmu-miR-205. To our knowledge, this is the first miRNA expression profiling study of external ear development in mammals. These data could set the basis to understand the implications of miRNAs in normal external ear development.
MicroRNAs (miRNAs)包括一类大约22个核苷酸调控的非编码rna,在不同的生物过程中发挥多种作用。最近的报道表明,哺乳动物的胚胎发育伴随着miRNA表达的动态变化;然而,没有关于mirna在外耳发育中的作用的信息。本研究的目的是确定小鼠外耳发育过程中mirna的阶段特异性表达,以鉴定潜在的相关mirna及其可能的靶点。使用Affymetrix GeneChip miRNA 3.0阵列获得胎鼠13.5 dpc和14.5 dpc时耳廓和背部皮肤组织的miRNA表达谱。对这两种组织的生物学副本进行了分析,每个副本来自一窝平均16个胎儿。使用Affymetrix的转录组分析控制台软件对结果进行分析,以鉴定差异表达的mirna。我们观察到40种miRNA的差异表达,包括一些被预测与外耳发育相关的靶基因,如mmu-miR-10a,一种已知调节Hoxa1 mRNA水平的miRNA,以及mmu-miR-200c和mmu-miR-205。据我们所知,这是首次研究哺乳动物外耳发育的miRNA表达谱。这些数据可以为理解mirna在正常外耳发育中的意义奠定基础。
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引用次数: 10
Visualization of primordial germ cells in the fertilized pelagic eggs of the barfin flounder Verasper moseri. 棘牙鲆远洋受精卵中原始生殖细胞的可视化。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.150008rg
R. Goto, Taiju Saito, Y. Kawakami, Tomoe Kitauchi, Misae Takagi, T. Todo, K. Arai, E. Yamaha
Primordial germ cells (PGCs) appear during early embryogenesis and differentiate into gametes through oogenesis or spermatogenesis. Teleost PGCs can be visualized by injecting RNA transcribed from the fusion product of a fluorescent protein gene attached to the 3' untranslated region (3'UTR) of zebrafish nanos3 (zf-nos3). Although this method has been widely applied to teleost PGCs, the visualization of PGCs in pelagic species that have eggs with a hard chorion is more problematic due to the technical difficulty of microinjection into their eggs. In this study, we developed a reliable method for microinjection of fertilized eggs in a pelagic species, the barfin flounder. Using a microneedle with a constriction "brake", we were able to introduce gfp-nos3 3'UTR mRNA into embryos and to determine the origin and migration route of PGCs. We also isolated the barfin flounder nos3 (bf-nos3) gene to compare its 3'UTR sequence with that of zebrafish. The 3'UTR of the bf-nos3 sequence was longer than that of zf-nos3. However, PGCs were also visualized after injection of gfp-bf-nos3 3'UTR mRNA both in zebrafish and barfin flounder. These results suggest that the function of nos3 is conserved between these species regardless of the sequence differences. The method developed here for labeling PGCs with gfp-nos3 mRNA will provide a means to study PGC development in the embryos of a wide range of marine fish species.
原始生殖细胞(PGCs)在胚胎发生早期出现,并通过卵发生或精子发生分化为配子。通过注射附着在斑马鱼nanos3 (zf-nos3) 3'非翻译区(3' utr)上的荧光蛋白基因融合产物转录的RNA,可以观察硬骨鱼PGCs。尽管该方法已广泛应用于硬骨鱼PGCs,但由于显微注射技术的困难,在具有硬绒毛膜的卵的远洋物种中可视化PGCs更加困难。在这项研究中,我们开发了一种可靠的显微注射受精卵的方法,在一个远洋物种,barfin比目鱼。使用带有收缩“制动器”的微针,我们能够将gfp-nos3 3'UTR mRNA引入胚胎,并确定PGCs的起源和迁移途径。我们还分离了barfin flder nos3 (bf-nos3)基因,并与斑马鱼的3'UTR序列进行了比较。bf-nos3序列的3′utr比zf-nos3长。然而,注射gfp-bf-nos3 3'UTR mRNA后,斑马鱼和斑马鱼的PGCs也可见。这些结果表明nos3的功能在这些物种之间是保守的,而不考虑序列差异。用gfp-nos3 mRNA标记PGCs的方法将为研究多种海洋鱼类胚胎中PGC的发育提供一种手段。
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引用次数: 16
Chicken primordial germ cell motility in response to stem cell factor sensing. 干细胞因子感应对鸡原始生殖细胞运动的影响。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.140287ct
Thanida Srihawong, T. Kuwana, K. Siripattarapravat, C. Tirawattanawanich
Avian primordial germ cells (PGCs) are destined to migrate a long distance from their extra embryonic region via the vascular system to the gonadal ridges where they form the germ cells. Although PGC migration is crucial for a genetic continuation to the next generation, the factors and mechanisms that control their migration remain largely unknown. In the present study the chemotactic effect of stem cell factor (SCF) was examined on chicken blood circulating PGCs (cPGC), employing 3D chemotaxis slides and time-lapsed imaging analyses as an in vitro study model. Upon in vitro exposure to an SCF gradient, 77.1% (54 out of 70) of cPGCs showed a clear response, of which 48.1% (26 out of 54) polarized with the consecutive formation of a persistent membrane protrusion and significant directional migration towards the gradient and the others showed transient membrane protrusions. In contrast, the controls and apparently SCF unresponsive cPGCs and c-kit-negative red blood cells (RBCs) showed only cytoplasmic cycling with random formations of membrane blebbing and no directional migration. Significant (p < 0.05) differences between the SCF-treated and control cPGCs and RBCs were found in the migration parameters of eccentricity, accumulated and Euclidean distances, and migration velocity. The SCF-treated PGCs also revealed a chemotactic response, as judged by their significant displacement of center of mass and Rayleigh test. Complete inhibition of all the SCF-induced responses in PGCs was found following pretreatment of the cPGCs with 10 µM of the c-kit inhibitor, STI57l, prior to SCF exposure. In addition, cPGCs were found to be positive for c-kit expression using a polyclonal goat anti-mouse c-kit primary antibody, suggesting that the cPGCs were capable of SCF sensing and the potential involvement of SCF/c-kit in the chemotactic migration. Therefore, SCF is suggested to function as a chemoattractant in the migration of chicken cPGC.
鸟类原始生殖细胞(PGCs)注定要通过血管系统从胚胎外区域迁移到性腺脊,在那里它们形成生殖细胞。尽管PGC迁移对于遗传延续到下一代至关重要,但控制其迁移的因素和机制在很大程度上仍然未知。本研究采用三维趋化载玻片和延时成像分析作为体外研究模型,探讨了干细胞因子(SCF)对鸡血液循环PGCs (cPGC)的趋化作用。在体外暴露于SCF梯度时,77.1%(54 / 70)的cpgc表现出明显的反应,其中48.1%(26 / 54)极化,连续形成持久的膜突出,并向梯度方向明显迁移,其他cpgc表现出短暂的膜突出。相比之下,对照组和明显无SCF反应的cpgc和c-kit阴性红细胞(rbc)仅显示细胞质循环,随机形成膜泡,无定向迁移。scf处理的cpgc和红细胞在偏心率、累积距离和欧氏距离、迁移速度等参数上与对照组有显著差异(p < 0.05)。经scf处理的PGCs也显示出趋化反应,通过质心的显著位移和瑞利试验来判断。在暴露于SCF之前,用10µM的c-kit抑制剂STI57l预处理cpgc后,发现PGCs中所有SCF诱导的反应都被完全抑制。此外,使用山羊抗小鼠c-kit一抗发现cPGCs的c-kit表达阳性,这表明cPGCs能够感知SCF,并且SCF/c-kit可能参与趋化迁移。因此,SCF可能是鸡cPGC迁移的化学引诱剂。
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引用次数: 11
CILP1 is dynamically expressed in the developing musculoskeletal system of the trout. CILP1在鳟鱼发育中的肌肉骨骼系统中动态表达。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.150136pr
C. Rallière, M. Frétaud, V. Thermes, P. Rescan
An in situ screen for genes expressed in the skeletal muscle of eyed-stage trout embryos led to the identification of a transcript encoding a polypeptide related to CILP1, a secreted glycoprotein present in the extracellular matrix. In situ hybridisation in developing trout embryos revealed that CILP1 expression was initially detected in fast muscle progenitors of the early somite. Later, CILP1 expression was down-regulated medio-laterally in differentiating fast muscle cells, to become finally restricted to the undifferentiated muscle progenitors forming the dermomyotome-like epithelium at the surface of the embryonic myotome. At the completion of somitogenesis, CILP1 expression was concentrated in the myoseptal/tendon cells that develop between adjacent myotomes but was excluded from the skeletogenic cells of the vertebral axis to which the most medial myoseptal/tendon cells attach. Overall, our work shows that muscle cells and myoseptal/tendon cells contribute dynamically and cooperatively to the production of CILP1 during ontogeny of the trout musculoskeletal system.
对眼期鳟鱼胚胎骨骼肌中表达的基因进行原位筛选,鉴定出编码与CILP1相关的多肽的转录本,CILP1是细胞外基质中分泌的糖蛋白。在发育中的鳟鱼胚胎中原位杂交发现,CILP1最初在早期体体的快肌祖细胞中被检测到。随后,在快速肌细胞分化过程中,CILP1的表达向中外侧下调,最终局限于未分化的肌祖细胞,形成胚胎肌瘤表面的皮细胞样上皮。在躯体发育完成时,CILP1的表达集中在相邻肌切块之间发育的肌间隔/肌腱细胞中,而在大多数内侧肌间隔/肌腱细胞附着的椎轴成骨细胞中则被排除在外。总的来说,我们的研究表明,在鳟鱼肌肉骨骼系统的个体发育过程中,肌肉细胞和肌隔/肌腱细胞对CILP1的产生做出了动态和合作的贡献。
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引用次数: 3
iTRAQ-based proteomic analysis of adaptive response in the regenerating limb of the Cynops orientalis newt. 基于itraq的东方蝾螈肢体再生适应性反应的蛋白质组学分析。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.150363cx
X. Geng, Jianlin Guo, Xiayan Zang, Jingyan Sun, Pengfei Li, Fu-chun Zhang, Cunshuan Xu
The newt has the powerful capacity to regenerate lost limbs following amputation, and represents an excellent model organism to study regenerative processes. However, the molecular basis of the adaptive response in the regenerating limb of the Chinese fire-bellied newt Cynops orientalis immediately after amputation remains unclear. To better understand the adaptive response immediately after limb amputation at the protein level, we used isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS methods to analyze changes in the proteome of the regenerating newt limb that occurred 2 h and 8 h after amputation. We identified 152 proteins with more than 1.5-fold change in expression compared to control. GO annotation analysis classified these proteins into several categories such as signaling, Ca(2+) binding and translocation, transcription and translation, immune response, cell death, cytoskeleton, metabolism, etc. Further ingenuity pathway analysis (IPA) showed that several signaling pathways were significantly changed at 2 h and 8 h after amputation, including EIF2 signaling, acute phase response signaling, tight junction signaling and calcium signaling, suggesting these pathways may be closely related to the adaptive response immediately after limb amputation. This work provides novel insights into understanding the molecular processes related to newt limb regeneration immediately after amputation, and a basis for further study of regenerative medicine.
蝾螈在截肢后具有强大的肢体再生能力,是研究再生过程的良好模式生物。然而,中国火腹蝾螈截肢后立即再生肢体适应性反应的分子基础尚不清楚。为了更好地了解断肢后在蛋白质水平上的适应性反应,我们采用等压标签相对和绝对定量(iTRAQ)结合LC-MS/MS方法分析断肢后2 h和8 h再生蝾螈肢体蛋白质组的变化。我们鉴定出152个蛋白与对照相比表达变化超过1.5倍。GO注释分析将这些蛋白分为信号转导、Ca(2+)结合与易位、转录与翻译、免疫应答、细胞死亡、细胞骨架、代谢等几类。进一步的匠心通路分析(ingenuity pathway analysis, IPA)显示,在截肢后2 h和8 h, EIF2信号、急性期反应信号、紧结信号和钙信号等信号通路发生了显著变化,提示这些信号通路可能与截肢后即刻的适应性反应密切相关。这项工作为理解蝾螈截肢后肢体再生的分子过程提供了新的见解,为再生医学的进一步研究奠定了基础。
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引用次数: 3
TRPM channels and magnesium in early embryonic development. TRPM通道和镁在早期胚胎发育中的作用。
Pub Date : 2015-01-01 DOI: 10.1387/ijdb.150196lr
Y. Komiya, L. Runnels
Magnesium (Mg(2+)) is the second most abundant cellular cation and is essential for all stages of life, from the early embryo to adult. Mg(2+) deficiency causes or contributes to many human diseases, including migraine headaches, Parkinson's disease, Alzheimer's disease, hypotension, type 2 diabetes mellitus and cardiac arrhythmias. Although the concentration of Mg(2+) in the extracellular environment can vary significantly, the total intracellular Mg(2+) concentration is actively maintained within a relatively narrow range (14 - 20 mM) via tight, yet poorly understood, regulation of intracellular Mg(2+)by Mg(2+) transporters and Mg(2+)-permeant ion channels. Recent studies have continued to add to the growing number of Mg(2+) transporters and ion channels involved in Mg(2+) homeostasis, including TRPM6 and TRPM7, members of the transient receptor potential (TRP) ion channel family. Mutations in TRPM6, including amino acid substitutions that prevent its heterooligomerization with TRPM7, occur in the rare autosomal-recessive disease hypomagnesemia with secondary hypocalcemia (HSH). Genetic ablation of either gene in mice results in early embryonic lethality, raising the question of whether these channels' capacity to mediate Mg(2+) influx plays an important role in embryonic development. Here we review what is known of the function of Mg(2+) in early development and summarize recent findings regarding the function of the TRPM6 and TRPM7 ion channels during embryogenesis.
镁(Mg(2+))是第二丰富的细胞阳离子,从早期胚胎到成年,在生命的各个阶段都是必不可少的。镁(2+)缺乏导致或促成了许多人类疾病,包括偏头痛、帕金森病、阿尔茨海默病、低血压、2型糖尿病和心律失常。尽管细胞外环境中Mg(2+)的浓度可以有很大的变化,但通过Mg(2+)转运体和Mg(2+)渗透离子通道对细胞内Mg(2+)的严格调控,胞内Mg(2+)的总浓度被积极地维持在一个相对狭窄的范围内(14 - 20 mM)。最近的研究继续增加了越来越多的Mg(2+)转运体和参与Mg(2+)稳态的离子通道,包括TRPM6和TRPM7,它们是瞬时受体电位(TRP)离子通道家族的成员。TRPM6的突变,包括阻止其与TRPM7异聚的氨基酸替换,发生在罕见的常染色体隐性疾病低镁血症伴继发性低钙血症(HSH)中。在小鼠中,任何一种基因的基因消融都会导致早期胚胎死亡,这就提出了这些通道介导Mg(2+)内流的能力是否在胚胎发育中起重要作用的问题。在这里,我们回顾了Mg(2+)在早期发育中的已知功能,并总结了最近关于TRPM6和TRPM7离子通道在胚胎发生过程中的功能的发现。
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引用次数: 37
Corrigendum: Hox collinearity - a new perspective (vol 55, pg 899, 2011) 勘误表:Hox共线性——一个新的视角(第55卷,第899页,2011)
Pub Date : 2012-04-24 DOI: 10.1387/IJDB.123515CO
A. Durston, H. Jansen, P. I. D. Rieden, M. Hooiveld
Hox collinearity is a spectacular phenomenon that has excited life scientists since its discovery in 1978. Two mechanisms have been proposed to explain the spatially sequential pattern of Hox gene expression in animal embryonic development: interactions among Hox genes, or the progressive opening of chromatin in the Hox clusters, from 3' to 5'. A review of the evidence across different species and developmental stages points to the universal involvement of trans-acting factors and cell-cell interactions. The evidence focuses attention on interactions between Hox genes and on the vertebrate somitogenesis clock. These novel conclusions open new perspectives for the field.
自1978年发现Hox共线性以来,它是一种壮观的现象,令生命科学家兴奋不已。动物胚胎发育中Hox基因表达的空间序列模式有两种解释机制:Hox基因之间的相互作用,或者Hox簇中染色质从3'到5'的逐渐开放。对不同物种和发育阶段的证据的回顾指出,反式作用因子和细胞-细胞相互作用的普遍参与。证据集中在Hox基因和脊椎动物体细胞发生时钟之间的相互作用。这些新颖的结论为该领域开辟了新的视角。
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引用次数: 1
CORRIGENDUM: Persistent expression of Twist1 in chondrocytes causes growth plate abnormalities and dwarfism in mice 更正:软骨细胞中Twist1的持续表达导致小鼠生长板异常和侏儒症
Pub Date : 2012-04-24 DOI: 10.1387/IJDB.123532CO
Rosa M. Guzzo, V. Andreeva, D. Spicer, M. Drissi
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引用次数: 0
期刊
The International journal of developmental biology
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