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Mild cryoinjury in zebrafish fin induces regenerative response without blastema formation.
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-02-01 DOI: 10.1111/dgd.12962
Takafumi Yoshida, Atsushi Kawakami

Previous studies have shown that tissue regeneration induces expression of genes that play important roles in regeneration. Recently, several studies have identified regeneration-response enhancers (RREs) that activate gene expression by tissue injury. Particularly, we showed that RREs contain two transcription factor-binding motifs: a bHLH transcription factor-binding motif, an E-box, and an AP-1/bZIP transcription factor-binding motif, a 12-O-Tetradecanoylphorbol 13-acetate response element (TRE). However, the triggers and subsequent signals generated by injury are still unclear. In this study, we analyzed RRE activation using various injury models. Although inter-ray incisions and skin exfoliation injuries did not activate RREs or regeneration genes, the fin puncture injury activated RREs and several regeneration-response genes. After fin puncture injury, msxc was activated only on the proximal side of the hole where blastema-like tissue was formed, whereas RREs, junbb, and fibronectin 1b (fn1b) were activated on both the proximal and distal sides, implying that activation of RREs, junbb, and fn1b is independent of blastema formation. Here, we also established a mild cryoinjury method. After this injury, transient vascular destruction, an increase in cell death, and an accumulation of myeloid cells were observed; however, no major morphological damage was observed. Importantly, msxc was not induced by cryoinjury, whereas fn1b, junbb, and 1.8 k RRE (-1.8 kb promoter of fn1b) were activated, suggesting that cryoinjury induces the responses of fn1b, junbb, and 1.8 k RRE without forming the blastema. Thus, our study shows that the cryoinjury model and the RRE transgenic (Tg) zebrafish may provide a useful platform for exploring injury signals.

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引用次数: 0
Understanding disorders of the human nervous system: How fish models reveal disease mechanisms from single molecules to behavior (part 2)
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-23 DOI: 10.1111/dgd.12951
Christina Lillesaar, William Norton, Daniel Liedtke, Sachiko Tsuda

The usefulness of zebrafish for understanding the human nervous system is exemplified by the articles in part 1. The virtual special issue part 2 not only covers more work using this well-established species, but also highlights that other fish species may serve as alternative or more appropriate models, due to unique biological or evolutionary characteristics, to explore genetic and molecular mechanisms of neurological and psychiatric disorders.

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引用次数: 0
Regeneration of Lumbriculus variegatus requires post-amputation production of reactive oxygen species. 畸形蚓的再生需要在截肢后产生活性氧。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-21 DOI: 10.1111/dgd.12961
Freya R Beinart, Kathy Gillen

Animals vary in their ability to replace body parts lost to injury, a phenomenon known as restorative regeneration. Uncovering conserved signaling steps required for regeneration may aid regenerative medicine. Reactive oxygen species (ROS) are necessary for proper regeneration in species across a wide range of taxa, but it is unknown whether ROS are essential for annelid regeneration. As annelids are a widely used and excellent model for regeneration, we sought to determine whether ROS play a role in the regeneration of the highly regenerative annelid, Lumbriculus variegatus. Using a ROS-sensitive fluorescent probe we observed ROS accumulation at the wound site within 15 min after amputation; this ROS burst lessened by 6 h post-amputation. Chemical inhibition of this ROS burst delayed regeneration, an impairment that was partially rescued with exogenous ROS. Our results suggest that similar to other animals, annelid regeneration depends upon ROS signaling, implying a phylogenetically ancient requirement for ROS in regeneration.

动物们替换受伤身体部位的能力各不相同,这种现象被称为恢复性再生。揭示再生所需的保守信号步骤可能有助于再生医学。活性氧(Reactive oxygen species, ROS)对于物种的正常再生是必需的,但对于环节动物的再生是否必需尚不清楚。由于环节动物是一种广泛使用的优秀的再生模型,我们试图确定ROS是否在高度再生的环节动物蓝斑虫(Lumbriculus variegatus)的再生中发挥作用。使用ROS敏感荧光探针,我们观察了截肢后15分钟内伤口部位的ROS积累;这种ROS爆发在截肢后6小时减弱。这种ROS爆发的化学抑制延迟了再生,这是外源性ROS部分修复的损伤。我们的研究结果表明,与其他动物类似,环节动物的再生依赖于ROS信号,这意味着在再生过程中对ROS有一个古老的系统发育要求。
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引用次数: 0
"Fly to New World": Meeting report of the 16th Japanese Drosophila research conference (JDRC16). “飞向新世界”:第16届日本果蝇研究大会(JDRC16)会议报告。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-18 DOI: 10.1111/dgd.12959
Haruka Yoshizawa, Erina Kuranaga

The 16th Japanese Drosophila Research Conference (JDRC16) was held at the Sendai International Center from September 17 to 19 2024. It had been 2 years since the last JDRC15 held in Nagoya. The conference brought together 231 researchers, including 22 researchers from overseas, creating a vibrant and diverse platform for scientific exchange. Prof. Shigeo Hayashi of RIKEN BDR delivered a keynote lecture, and his groundbreaking ideas and research captivated the audience. Over the 3 days, the conference featured 53 oral presentations across 11 sessions and 2 special sessions, as well as 128 poster presentations, all of which fostered stimulating discussions and the exchange of innovative ideas. The reception provided an additional opportunity for researchers to engage in meaningful dialogue while enjoying Sendai's renowned specialties. Held under clear autumn skies in a great nature along the river, this conference painted a beautiful contrast to the heated discussions in the venue. Consequently, this conference fully contributed to the mission proffered by Prof. Hayashi, "Fly to New World," expanding the insights gained from flies into new and unexplored scientific areas.

第16届日本果蝇研究大会(JDRC16)于2024年9月17日至19日在仙台国际中心召开。上一次在名古屋举行的jdrc - 15会议已经过去两年了。会议汇集了231名研究人员,其中包括22名来自海外的研究人员,为科学交流创造了一个充满活力和多样化的平台。RIKEN BDR的Shigeo Hayashi教授发表了主题演讲,他开创性的想法和研究吸引了观众。在为期三天的会议中,共有11场会议和2场特别会议共53场口头报告,以及128场海报展示,所有这些都促进了热烈的讨论和创新思想的交流。招待会为研究人员提供了额外的机会,在享受仙台著名特产的同时进行有意义的对话。这次会议在秋高气爽的天气下,在河边的美景中举行,与会场内激烈的讨论形成了鲜明的对比。因此,这次会议对Hayashi教授提出的“飞向新世界”的使命做出了充分的贡献,将从苍蝇身上获得的见解扩展到新的和未开发的科学领域。
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引用次数: 0
The transmembrane protein TMEM196 controls cell proliferation and determines the floor plate cell lineage. 跨膜蛋白TMEM196控制细胞增殖并决定底板细胞谱系。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-18 DOI: 10.1111/dgd.12960
Yumi Matsumoto, Seiichi Tamaru, Xing Chen, Takuma Shinozuka, Yuichi Sakumura, Noriaki Sasai

The neural tube, the embryonic precursor to the vertebrate central nervous system, comprises distinct progenitor and neuronal domains, each with specific proliferation programs. In this study, we identified TMEM196, a novel transmembrane protein that plays a crucial role in regulating cell proliferation in the floor plate in chick embryos. TMEM196 is expressed in the floor plate, and its overexpression leads to reduced cell proliferation without affecting the pattern formation of the neural tube. We also established the floor plate differentiation protocol of the mouse embryonic stem cells, and analyzed the function of TMEM196 with this system. Mutating the Tmem196 gene does not alter cell division and overall differentiation remains unchanged within the neural cells. However, TMEM196 inhibits Wnt signaling, and Tmem196 mutant cells exhibit aberrant paraxial mesoderm differentiation, suggesting that TMEM196 selects the floor plate cell fate at the binary decision of the neuromesodermal cells. These findings highlight TMEM196 as a key regulator of both cell proliferation and floor plate determination, contributing to proper regionalization during embryogenesis.

神经管是脊椎动物中枢神经系统的胚胎前体,由不同的祖细胞和神经元结构域组成,每个结构域都有特定的增殖程序。在这项研究中,我们发现了一种新的跨膜蛋白TMEM196,它在调节鸡胚胎底板细胞增殖中起着至关重要的作用。TMEM196在底板中表达,其过表达导致细胞增殖减少,但不影响神经管图案的形成。我们还建立了小鼠胚胎干细胞的底板分化方案,并利用该系统分析了TMEM196的功能。突变Tmem196基因不会改变细胞分裂,神经细胞内的整体分化保持不变。然而,TMEM196抑制Wnt信号,TMEM196突变细胞表现出异常的近轴中胚层分化,表明TMEM196在神经中胚层细胞的二元决策中选择了底板细胞的命运。这些发现强调了TMEM196是细胞增殖和底板决定的关键调节因子,有助于胚胎发生过程中的适当区域化。
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引用次数: 0
Early development of the calcified exoskeleton of the polyplacophoran mollusk, with insight into the evolutionary history of shell plates and spicules. 多placophoran软体动物钙化外骨骼的早期发育,并深入了解壳板和针状体的进化史。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-10 DOI: 10.1111/dgd.12956
Hiroki Yoshikawa, Yoshiaki Morino, Hiroshi Wada

Recent molecular phylogenetic studies have raised two questions about the evolutionary history of the calcified exoskeleton of mollusks. The first question concerns the homology of the two types of skeleton: whether spicules and shell plates share an evolutionary origin. The second question is the homology of the shell plates between chitons and other mollusks, including gastropods and bivalves. To gain insight into these questions, we examined the early development of shell plates and spicules in chitons. We identified several developmental genes that are involved in both shell plates and spicules, suggesting that spicules and shell plates share a common evolutionary origin. We also found that subpopulations of the dorsal shell field (the ridge and the plate field) have specific gene expression profiles. The differential gene expression of the ridge and plate field is not identical to the profiles of the zones of the gastropod shell field. This observation may suggest an independent evolutionary origin of the shell plates in chitons and gastropods.

最近的分子系统发育研究对软体动物钙化外骨骼的进化史提出了两个问题。第一个问题涉及两种骨骼的同源性:针状体和壳板是否有共同的进化起源。第二个问题是石鳖和其他软体动物(包括腹足类动物和双壳类动物)的壳板的同源性。为了深入了解这些问题,我们研究了石鳖壳板和针状体的早期发育。我们发现了几个涉及壳板和针状体的发育基因,这表明针状体和壳板具有共同的进化起源。我们还发现背壳场(脊和板场)的亚群具有特定的基因表达谱。脊区和板区基因的差异表达与腹足类壳区基因的差异表达不完全相同。这一观察结果可能表明石鳖和腹足类动物的壳板具有独立的进化起源。
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引用次数: 0
Sonic Hedgehog signaling regulates the optimal differentiation pace from early-stage mesoderm to cardiogenic mesoderm in mice. Sonic Hedgehog信号调控小鼠早期中胚层向心源性中胚层的最佳分化速度。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-09 DOI: 10.1111/dgd.12955
Satoshi Inoue, Moe Nosetani, Yoshiro Nakajima, Shinichiro Sakaki, Hiroki Kato, Rie Saba, Naoki Takeshita, Kosuke Nishikawa, Atsuko Ueyama, Kazuhiko Matsuo, Masaki Shigeta, Daisuke Kobayashi, Tomoko Iehara, Kenta Yashiro

Sonic Hedgehog (Shh), encoding an extracellular signaling molecule, is vital for heart development. Shh null mutants show congenital heart disease due to left-right asymmetry defects stemming from functional anomaly in the midline structure in mice. Shh signaling is also known to affect cardiomyocyte differentiation, endocardium development, and heart morphogenesis, particularly in second heart field (SHF) cardiac progenitor cells that contribute to the right ventricle, outflow tract, and parts of the atrium. Despite extensive studies, our understanding remains incomplete. Notably, Shh signaling is suggested to promote cardiac differentiation, while paradoxically preventing premature differentiation of SHF progenitors. In this study, we elucidate the role of Shh signaling in the earliest phase of cardiac differentiation. Our meta-analysis of single-cell RNA sequencing suggests that cardiogenic nascent mesoderm cells expressing the bHLH transcription factor Mesp1 interact with axial mesoderm via Hh signaling. Activation of Hh signaling using a Smoothened agonist delayed or suppressed the differentiation of primitive streak cells expressing T-box transcription factor T to Mesp1+ nascent mesoderm cells both in vitro and ex vivo. Conversely, inhibition of Hh signaling by cyclopamine facilitated cardiac differentiation. The reduction of Eomes, an inducer of Mesp1, by Hh signaling appears to be the underlying mechanism of this phenomenon. Our data suggest that SHH secreted from axial mesoderm inhibits premature differentiation of T+ cells to Mesp1+ nascent mesoderm cells, thereby regulating the pace of cardiac differentiation. These findings enhance our comprehension of Shh signaling in cardiac development, underscoring its crucial role in early cardiac differentiation.

Sonic Hedgehog (Shh)编码细胞外信号分子,对心脏发育至关重要。Shh缺失突变体表现出先天性心脏病,这是由于小鼠中线结构功能异常引起的左右不对称缺陷。众所周知,Shh信号也会影响心肌细胞分化、心内膜发育和心脏形态发生,特别是在第二心田(SHF)心脏祖细胞中,这些细胞对右心室、流出道和部分心房起作用。尽管进行了广泛的研究,我们的理解仍然不完整。值得注意的是,Shh信号被认为可以促进心脏分化,同时矛盾地阻止SHF祖细胞的过早分化。在这项研究中,我们阐明了Shh信号在心脏分化的早期阶段的作用。我们对单细胞RNA测序的meta分析表明,表达bHLH转录因子Mesp1的心源性新生中胚层细胞通过Hh信号与轴向中胚层相互作用。在体外和离体实验中,使用Smoothened激动剂激活Hh信号可以延迟或抑制表达T-box转录因子T的原始条纹细胞向Mesp1+新生中胚层细胞的分化。相反,环巴胺对Hh信号的抑制促进了心脏分化。通过Hh信号传导减少Eomes (Mesp1的诱导剂)似乎是这一现象的潜在机制。我们的数据表明,轴向中胚层分泌的SHH抑制T+细胞向Mesp1+新生中胚层细胞的过早分化,从而调节心脏分化的速度。这些发现增强了我们对Shh信号在心脏发育中的理解,强调了其在早期心脏分化中的关键作用。
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引用次数: 0
A comparative approach to the microstructure in the carpus and tarsus in anurans. anurans腕骨和跗骨微观结构的比较研究。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-08 DOI: 10.1111/dgd.12957
Marissa Fabrezi, Julio César Cruz

Despite the significant literature about morphological features of limb skeletons involved in tetrapod limb evolution, some questions about carpal and tarsal elements remain. In anurans, the ecomorphological and biomechanical approaches studied long hind limbs (to jump) and forelimbs (to land) and emphasized the role of the long bones in locomotion but disregarded what happens with the nodular elements of the carpus and tarsus. Here, we present a comparative study of nodular elements of the carpus and tarsus in anurans based on whole-mount specimens stained with Alcian Blue (cartilage) and Alizarin Red S (bone and calcified cartilage). The sample comprises 113 species belonging to 33 anuran families and postmetamorphic series in selected species. Further, we analyze the histology of the carpus and tarsus in individuals of nine species. In most anurans, the carpal and tarsal elements are cartilaginous in adult stages. The cartilaginous matrix may present different degrees of calcification. Few taxa present truly ossified carpals and tarsals with marrow cavity, blood cells, and hematopoietic cells. Interpretation of the interspecific variation in the carpus and tarsus skeletons on the most recent anuran phylogeny suggests that the delayed ossification of carpals and tarsals has evolved in derived lineages (e.g. Pelobatoidea and Neobatrachia).

尽管关于四肢骨骼在四足动物肢体进化过程中的形态学特征的研究已有大量文献,但关于腕骨和跗骨的研究仍存在一些问题。在无尾动物中,生态形态学和生物力学方法研究了长后肢(用于跳跃)和前肢(用于着陆),并强调了长骨在运动中的作用,但忽略了腕骨和跗骨结节元素的作用。在此,我们基于阿利新蓝(软骨)和茜素红S(骨和钙化软骨)染色的整座标本,对anurans腕骨和跗骨的结节元素进行了比较研究。样本包括33个菊科113种,部分属后变质系。此外,我们分析了9种个体的腕骨和跗骨的组织学。在大多数无脊椎动物中,腕骨和跗骨在成体阶段是软骨。软骨基质可呈现不同程度的钙化。少数分类群呈现真正骨化的腕骨和跗骨,有骨髓腔、血细胞和造血细胞。对腕骨和跗骨骨骼在最新无脊椎动物系统发育中的种间变异的解释表明,腕骨和跗骨的延迟骨化已经在衍生谱系中进化(例如Pelobatoidea和Neobatrachia)。
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引用次数: 0
Effect of Cdk1 gene disruption on cell cycle progression in newt cells. Cdk1基因破坏对蝾螈细胞周期进程的影响。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-08 DOI: 10.1111/dgd.12958
Yuta Nakao, Kazuko Okamoto, Ichiro Tazawa, Tatsuro Nishijima, Nobuaki Furuno, Tetsushi Sakuma, Takashi Yamamoto, Takashi Takeuchi, Toshinori Hayashi

Cyclin-dependent kinases (CDKs) are key regulators of cell cycle progression, in conjunction with cyclins. The cyclin-CDK system is highly conserved among eukaryotes, and CDK1 is considered essential for progression through the M phase. However, the extent to which cell cycle progression depends on CDK1 varies between cell types. Therefore, a range of cell types must be analyzed to comprehensively elucidate the role of CDK1. Cdk1-knockout mice exhibit lethality at an early developmental stage, specifically before the differentiation of various cell types. The aim of the present study was to characterize the effects of CDK1 deficiency in amphibian newts. Cdk1 was disrupted by injecting fertilized newt eggs with CRISPR/Cas9, and the resulting effects on embryonic development and cell proliferation were then evaluated. In both wild-type and Cdk1-crispant newt embryos, CDK1 protein was either stored in the egg until late embryogenesis or potentially derived from maternal mRNA, which may also be stored during this period. The embryos survived to the hatching stage, during which the cells responsible for forming the basic organs differentiated. To further characterize the long-term effects of Cdk1 knockout, parabiosis experiments were conducted using wild-type embryos and Cdk1 crispants. The results suggested that an endocycle occurred in the crispant larvae, as evidenced by increases in the size of several types of cells. It is anticipated that studies using newts will provide further insights into the role of Cdk1 in regulating the cell cycle.

细胞周期蛋白依赖性激酶(CDKs)与细胞周期蛋白一起是细胞周期进程的关键调节因子。周期蛋白- cdk系统在真核生物中是高度保守的,CDK1被认为是通过M期进展所必需的。然而,细胞周期进程依赖CDK1的程度因细胞类型而异。因此,必须对一系列细胞类型进行分析,以全面阐明CDK1的作用。cdk1敲除小鼠在早期发育阶段表现出致命性,特别是在各种细胞类型分化之前。本研究的目的是表征CDK1缺乏对两栖蝾螈的影响。通过向受精卵注射CRISPR/Cas9来破坏Cdk1,然后评估其对胚胎发育和细胞增殖的影响。在野生型和CDK1脆型蝾螈胚胎中,CDK1蛋白要么储存在卵子中直到胚胎发生晚期,要么可能来源于母体mRNA,后者也可能在此期间储存。胚胎存活到孵化阶段,在此期间,负责形成基本器官的细胞分化。为了进一步表征Cdk1基因敲除的长期影响,我们使用野生型胚胎和Cdk1 crispants进行了异种共生实验。结果表明,在脆嫩的幼虫中出现了一个内循环,这可以从几种类型细胞的大小增加中得到证明。预计对蝾螈的研究将进一步深入了解Cdk1在调节细胞周期中的作用。
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引用次数: 0
Meeting report about self-organization in biology: Freiburg Spemann-Mangold Centennial Symposium. 生物学中自组织的会议报告:弗莱堡Spemann-Mangold百年学术研讨会。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2025-01-08 DOI: 10.1111/dgd.12954
Satoshi Kuwana, Yuuri Yasuoka

From September 16 to 19, 2024, an international symposium to celebrate the centennial of the discovery of the gastrula organizer by Hans Spemann and Hilde Mangold, was held at the University of Freiburg, Germany, where they studied embryology. There were 41 plenary lectures, 11 short talks, and 182 poster presentations, with more than 300 participants from 23 countries. The symposium covered research topics broadly related to developmental, cell, genome, and evolutionary biology, mainly focused on early animal development. In addition to in vivo studies on topics such as gastrulation, embryonic patterning, cell polarity, and morphogenesis, recent studies using gastruloids and organoids, which recapitulate embryogenesis and organogenesis in in vitro cell culture, were also presented at this symposium, entitled Self-Organization in Biology. Most of the reported studies used vertebrate models such as mice, frogs, and zebrafish; however, evolutionary studies involving invertebrate and plant models were also presented. Presentations employing traditional methods such as cell transplantation and phenotype screening, and state-of-the-art technologies such as single-cell omics, high-resolution imaging, and computational analysis showed that experimental embryology has a long history, to which studies of the organizer have contributed significantly. Here we discuss memorable aspects of the symposium in the hope that this report will encourage young scientists to actively participate in face-to-face international conferences.

2024年9月16日至19日,在德国弗莱堡大学举行了一次国际研讨会,以庆祝汉斯·斯皮曼和希尔德·曼戈尔德发现原肠菌100周年,他们曾在那里研究胚胎学。共有来自23个国家的300多人参加了41场全体演讲、11场简短演讲和182场海报展示。研讨会涵盖了与发育、细胞、基因组和进化生物学广泛相关的研究主题,主要集中在早期动物发育方面。除了关于原肠胚形成、胚胎模式、细胞极性和形态发生等主题的体内研究外,最近关于在体外细胞培养中概括胚胎发生和器官发生的类原肠和类器官的研究也在本次题为“生物学中的自组织”的研讨会上进行了介绍。大多数报道的研究使用脊椎动物模型,如小鼠、青蛙和斑马鱼;然而,涉及无脊椎动物和植物模型的进化研究也被提出。采用细胞移植和表型筛选等传统方法以及单细胞组学、高分辨率成像和计算分析等最新技术的报告表明,实验胚胎学具有悠久的历史,组织者的研究对此做出了重大贡献。在这里,我们讨论研讨会中值得纪念的方面,希望这篇报告能鼓励年轻科学家积极参加面对面的国际会议。
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引用次数: 0
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Development Growth & Differentiation
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