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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
Novel function of Hox13 in regulating outgrowth of the newt hindlimb bud through interaction with Fgf10 and Tbx4 Hox13通过与Fgf10和Tbx4相互作用调控蝾螈后肢芽生长的新功能。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2024-12-26 DOI: 10.1111/dgd.12952
Sayo Tozawa, Haruka Matsubara, Fumina Minamitani, Yasuhiro Kamei, Misako Saida, Momoko Asao, Ken-ichi T. Suzuki, Masatoshi Matsunami, Shuji Shigenobu, Toshinori Hayashi, Gembu Abe, Takashi Takeuchi

5′Hox genes regulate pattern formation along the axes of the limb. Previously, we showed that Hoxa13/Hoxd13 double-mutant newts lacked all digits of the forelimbs during development and regeneration, showing that newt Hox13 is necessary for digit formation in development and regeneration. In addition, we found another unique phenotype. Some of the Hox13 crispant newts showed hindlimb defects, in which whole or almost whole hindlimbs were lost, suggesting a novel function of Hox13 in limb development. Using germline mutants, we showed that mutation in Hox13 led to hindlimb defects. The limb buds of Hox13 crispants formed, however, did not show outgrowth. Expression of Fgf10 and Tbx4, which are involved in limb outgrowth, decreased in the hindlimb buds of Hox13 crispants. In addition, hindlimb defects were observed in both Fgf10 and Tbx4 crispant newts. Finally, Fgf10 and Tbx4 interacted with Hox13 genetically. Our results revealed a novel function of Hox13 in regulating the outgrowth of the newt hindlimb bud through interaction with Fgf10 and Tbx4.

5'Hox基因调节沿肢轴的模式形成。之前,我们发现Hoxa13/Hoxd13双突变蝾螈在发育和再生过程中缺少前肢的所有趾,这表明蝾螈在发育和再生过程中趾的形成是必需的。此外,我们还发现了另一种独特的表型。一些Hox13脆嫩蝾螈表现出后肢缺陷,其中整个或几乎整个后肢缺失,这表明Hox13在肢体发育中具有新的功能。利用种系突变体,我们发现Hox13突变导致后肢缺陷。然而,Hox13 crispants形成的肢芽没有显示出生长。参与肢体生长的Fgf10和Tbx4在Hox13 crispants后肢芽中的表达降低。此外,在Fgf10和Tbx4脆嫩蝾螈中均观察到后肢缺陷。最后,Fgf10和Tbx4与Hox13基因互作。我们的研究结果揭示了Hox13通过与Fgf10和Tbx4相互作用调节蝾螈后肢芽生长的新功能。
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引用次数: 0
The polycomb protein complex interacts with GATA-6/PPARα to inhibit α-MHC expression 多梳蛋白复合物与GATA-6/PPARα相互作用抑制α-MHC表达。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2024-12-26 DOI: 10.1111/dgd.12953
Fei-Fei Dai, Jing Chen, Zhen Ma, Ming-Hui Yang, Tong Sun, Juan Ma, Meng-Jiao Zhou, Zhi-Ru Wei, Yunzeng Zou, Shoutao Zhang, Ming-Xi Zang

Transcription factors collaborate with epigenetic regulatory factors to orchestrate cardiac differentiation for heart development, but the underlying mechanism is not fully understood. Here, we report that GATA-6 induces cardiac differentiation but peroxisome proliferator-activated receptor α (PPARα) reverses GATA-6-induced cardiac differentiation, possibly because GATA-6/PPARα recruits the polycomb protein complex containing EZH2/Ring1b/BMI1 to the promoter of the cardiac-specific α-myosin heavy chain (α-MHC) gene and suppresses α-MHC expression, which ultimately inhibits cardiac differentiation. Furthermore, Ring1b ubiquitylates PPARα and GATA-6. By overexpression and knockout of EZH2/BMI1, it was demonstrated that the polycomb protein complex inhibits cardiac differentiation induced by GATA-6 and PPARα. Together, our results demonstrate that the polycomb protein complex interacts with GATA-6/PPARα to inhibit cardiac differentiation, a finding that could facilitate the development of new therapies for congenital heart disease.

转录因子与表观遗传调控因子协同协调心脏分化以促进心脏发育,但其潜在机制尚不完全清楚。在这里,我们报道了GATA-6诱导心脏分化,但过氧化物酶体增殖物激活受体α (PPARα)逆转了GATA-6诱导的心脏分化,可能是因为GATA-6/PPARα将含有EZH2/Ring1b/BMI1的多梳子蛋白复合物募集到心脏特异性α-肌球蛋白重链(α- mhc)基因的启动子上,抑制α- mhc的表达,最终抑制心脏分化。此外,Ring1b泛素化PPARα和GATA-6。通过过表达和敲除EZH2/BMI1,证实了多梳蛋白复合物抑制GATA-6和PPARα诱导的心脏分化。总之,我们的研究结果表明,多梳蛋白复合物与GATA-6/PPARα相互作用抑制心脏分化,这一发现可能促进先天性心脏病新疗法的发展。
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引用次数: 0
Meeting report: Society for Developmental Biology 83rd annual meeting 会议报告:发育生物学学会第83届年会。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2024-12-03 DOI: 10.1111/dgd.12950
Shunsuke Yaguchi
<p>The 83rd Annual Meeting of the Society for Developmental Biology (SDB) (https://www.sdbonline.org/2024mtg) was held in Atlanta, where the Japanese Society of Developmental Biologists (JSDB) participated as the guest society. To promote societal interactions, three researchers from Japan and gave invited talks in the SDB meeting as representatives of JSDB. On the other hand, four researchers from SDB gave invited talks and also participated in the Diversity Committee's Luncheon Seminar that I organized in the JSDB meeting (https://pub.confit.atlas.jp/en/event/jsdb2024). These scientific exchanges are aimed at fostering collaboration between developmental biologists in the United States and Japan and promoting mutual research advancements. The hope is that these initiatives will continue to build a long-term cooperative framework between both parties. Notably, this year, SDB President Dr. Ken Cho attended the JSDB meeting in Kyoto and played a significant role in facilitating our visit, underscoring the importance of mutual exchange.</p><p>I had the privilege of attending the Society for Developmental Biology (SDB) Annual Meeting for the first time in 2024. The 2024 SDB Annual Meeting took place at the Signia by Hilton, a newly opened venue in downtown Atlanta, Georgia (Figure 1). The hotel location offers excellent access to key landmarks such as the Mercedes-Benz Stadium and the Georgia World Congress Center. The city of Atlanta, with a population of approximately 500,000 as of 2022, is one of the major urban centers of the southern United States and is notable for hosting the 1996 Summer Olympics for our generations. The hotel provided an ideal setting for networking and engaging with colleagues, which complemented the productive scientific sessions held throughout the meeting.</p><p>The SDB meeting delivered a broad range of topics in developmental biology, and the sessions offered excellent opportunities for exchanging ideas and discussing the latest research advances. As a participant, I found the meeting to be highly valuable for staying updated with cutting-edge discoveries and methodologies, as well as for establishing new collaborations within the developmental biology community.</p><p>The scheduling format included a Presidential Symposium in the evening of day 1, with concurrent sessions and symposia in the morning, and plenary sessions after dinner. The invited speakers delivered outstanding presentations, both in terms of research content and presentation skills. The presentation by Dr. Zeba Wunderlich and her team from Boston University explored the functional significance of shadow enhancers, a critical yet enigmatic element in gene regulation. Using Drosophila embryos, they demonstrated how shadow enhancers, which bind distinct sets of transcription factors, ensure robust gene expression even under stress conditions. A particularly surprising finding came from their experiments with “squish” enhancers, where the endogenous DNA b
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引用次数: 0
Quantitative in toto live imaging analysis of apical nuclear migration in the mouse telencephalic neuroepithelium 小鼠端脑神经上皮细胞顶端核迁移的定量全图活体成像分析。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2024-11-26 DOI: 10.1111/dgd.12949
Tsukasa Shimamura, Takaki Miyata

In the embryonic neuroepithelium (NE), neural progenitor cells undergo cell cycle-dependent interkinetic nuclear migration (IKNM) along the apicobasal axis. Extensive IKNM supports increasing cell production rates per unit apical surface, as typically observed in the mammalian telencephalic NE. Apical nucleokinesis during the G2 phase is an essential premitotic event, but its occurrence has not yet been quantitatively analyzed at a large 3D-scale with sufficient spatiotemporal resolution. Here, we comprehensively analyzed apically migrating nuclei/somata in reference to their surroundings from embryonic day (E)11 to E13 in the mouse telencephalon. The velocity of apical nucleokinesis decreased, with more frequent nuclear pausing occurring at E12 and E13, whereas the nuclear density in the middle NE zone (20–40-μm deep) increased. This result, together with the results of Shh-mediated overproliferation experiments in which the nuclear density was increased in vivo at E11, suggests that apical nucleokinesis is physically influenced by the surrounding nuclei. Mean square displacement analysis for nuclei being passed by the apically migrating nuclei via horizontal sectioning in toto-recorded movies revealed that the “tissue fluidity” or physical permissiveness of the NE to apical nucleokinesis gradually decreased (E11 > E12 > E13). To further investigate the spatial relationship between preexisting mitoses and subsequent premitotic apical nucleokinesis, the horizontal distribution of mitoses was cumulatively (~3 hr) analyzed under in toto monitoring. The four-dimensional cumulative apical mitoses presented a “random”, not “clustered” or “regular”, distribution pattern throughout the period examined. These methodologies provide a basis for future comparative studies of interspecies differences.

在胚胎神经上皮(NE)中,神经祖细胞会沿着尖基轴进行细胞周期依赖性动核间迁移(IKNM)。广泛的IKNM支持每单位顶端表面的细胞生成率不断提高,这在哺乳动物端脑NE中是典型的现象。G2 期的顶端核运动是一个重要的减数分裂前事件,但其发生尚未在大三维尺度上以足够的时空分辨率进行定量分析。在这里,我们参照小鼠端脑从胚胎第 11 天到第 13 天的周围环境,全面分析了顶端迁移的细胞核/肿块。顶端核运动的速度下降,在 E12 和 E13 出现更频繁的核停顿,而中间 NE 区(20-40μm 深)的核密度增加。这一结果连同 Shh 介导的过度增殖实验结果(其中 E11 阶段体内核密度增加)表明,顶端核运动受到周围细胞核的物理影响。在全图记录的影片中,通过水平切片对被顶端迁移的细胞核穿过的细胞核进行均方位移分析,发现东北核对顶端核运动的 "组织流动性 "或物理允许性逐渐降低(E11 > E12 > E13)。为了进一步研究前期有丝分裂和随后的凋亡前顶端核动之间的空间关系,在整体监测下对有丝分裂的水平分布进行了累积分析(约 3 小时)。在整个研究期间,四维累积的顶端有丝分裂呈 "随机 "分布模式,而非 "集群 "或 "规则 "分布模式。这些方法为今后比较研究种间差异提供了基础。
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引用次数: 0
Labeling and sorting of avian primordial germ cells utilizing Lycopersicon Esculentum lectin 利用Lycopersicon Esculentum凝集素对禽类原始生殖细胞进行标记和分类。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2024-11-09 DOI: 10.1111/dgd.12948
Hiroko Iikawa, Aika Nishina, Mizuki Morita, Yuji Atsuta, Yoshiki Hayashi, Daisuke Saito

Avian species are essential resources for human society, with their preservation and utilization heavily dependent on primordial germ cells (PGCs). However, efficient methods for isolating live PGCs from embryos remain elusive in avian species beyond chickens, and even in chickens, existing techniques have shown limited efficiency. In this study, we present a rapid, simple, and cost-effective method for labeling and sorting circulating-stage PGCs across various avian species, including Carinatae and Ratitae, using Lycopersicon Esculentum (Tomato) lectin (LEL). Notably, this method demonstrates high sorting efficiency by identifying a wide range of PGC subtypes while preserving the proliferative and migratory potential of chicken PGCs. This approach is anticipated to significantly contribute to the conservation, research, and agricultural industries related to avian species globally.

禽类是人类社会的重要资源,其保存和利用在很大程度上依赖于原始生殖细胞(PGCs)。然而,在鸡以外的禽类物种中,从胚胎中分离活的 PGCs 的有效方法仍然难以找到,即使在鸡中,现有技术也显示出有限的效率。在本研究中,我们提出了一种快速、简单且经济有效的方法,利用番茄凝集素(LEL)对不同禽类物种(包括鲤科和鼠科)的循环期 PGCs 进行标记和分拣。值得注意的是,这种方法既能识别多种 PGC 亚型,又能保留鸡 PGC 的增殖和迁移潜能,因此具有很高的分拣效率。预计这种方法将为全球禽类物种的保护、研究和农业产业做出重大贡献。
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引用次数: 0
Transition from fetal to postnatal state in the heart: Crosstalk between metabolism and regeneration 心脏从胎儿状态向出生后状态的过渡:新陈代谢与再生之间的相互影响
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2024-10-27 DOI: 10.1111/dgd.12947
Tai Sada, Wataru Kimura

Cardiovascular disease is the leading cause of mortality worldwide. Myocardial injury resulting from ischemia can be fatal because of the limited regenerative capacity of adult myocardium. Mammalian cardiomyocytes rapidly lose their proliferative capacities, with only a small fraction of adult myocardium remaining proliferative, which is insufficient to support post-injury recovery. Recent investigations have revealed that this decline in myocardial proliferative capacity is closely linked to perinatal metabolic shifts. Predominantly glycolytic fetal myocardial metabolism transitions towards mitochondrial fatty acid oxidation postnatally, which not only enables efficient production of ATP but also causes a dramatic reduction in cardiomyocyte proliferative capacity. Extensive research has elucidated the mechanisms behind this metabolic shift, as well as methods to modulate these metabolic pathways. Some of these methods have been successfully applied to enhance metabolic reprogramming and myocardial regeneration. This review discusses recently acquired insights into the interplay between metabolism and myocardial proliferation, emphasizing postnatal metabolic transitions.

心血管疾病是导致全球死亡的主要原因。由于成人心肌的再生能力有限,缺血导致的心肌损伤可能是致命的。哺乳动物的心肌细胞会迅速失去增殖能力,只有一小部分成人心肌仍具有增殖能力,这不足以支持损伤后的恢复。最近的研究发现,心肌增殖能力的下降与围产期代谢转变密切相关。以糖酵解为主的胎儿心肌代谢在出生后向线粒体脂肪酸氧化过渡,这不仅能有效产生 ATP,还能导致心肌细胞增殖能力急剧下降。大量研究已经阐明了这种代谢转变背后的机制,以及调节这些代谢途径的方法。其中一些方法已成功应用于加强代谢重编程和心肌再生。本综述将讨论最近获得的有关新陈代谢与心肌增殖之间相互作用的见解,重点是出生后的新陈代谢转变。
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引用次数: 0
Mitochondrial DNA replication is essential for neurogenesis but not gliogenesis in fetal neural stem cells 线粒体DNA复制对胎儿神经干细胞的神经发生(neurogenesis)至关重要,但对神经胶质细胞的发生(gliogenesis)并不重要。
IF 1.7 4区 生物学 Q4 CELL BIOLOGY Pub Date : 2024-10-22 DOI: 10.1111/dgd.12946
Meri Walter-Manucharyan, Melanie Martin, Julia Pfützner, Franz Markert, Gerhard Rödel, Andreas Deussen, Andreas Hermann, Alexander Storch

Mitochondria are unique organelles that have their own genome (mtDNA) and perform various pivotal functions within a cell. Recently, evidence has highlighted the role of mitochondria in the process of stem cell differentiation, including differentiation of neural stem cells (NSCs). Here we studied the importance of mtDNA function in the early differentiation process of NSCs in two cell culture models: the CGR8-NS cell line that was derived from embryonic stem cells by a lineage selection technique, and primary NSCs that were isolated from embryonic day 14 mouse fetal forebrain. We detected a dramatic increase in mtDNA content upon NSC differentiation to adapt their mtDNA levels to their differentiated state, which was not accompanied by changes in mitochondrial transcription factor A expression. As chemical mtDNA depletion by ethidium bromide failed to generate living ρ° cell lines from both NSC types, we used inhibition of mtDNA polymerase-γ by 2′-3′-dideoxycytidine to reduce mtDNA replication and subsequently cellular mtDNA content. Inhibition of mtDNA replication upon NSC differentiation reduced neurogenesis but not gliogenesis. The mtDNA depletion did not change energy production/consumption or cellular reactive oxygen species (ROS) content in the NSC model used. In conclusion, mtDNA replication is essential for neurogenesis but not gliogenesis in fetal NSCs through as yet unknown mechanisms, which, however, are largely independent of energy/ROS metabolism.

线粒体是一种独特的细胞器,拥有自己的基因组(mtDNA),在细胞内发挥着各种关键功能。最近,有证据强调了线粒体在干细胞分化过程中的作用,包括神经干细胞(NSCs)的分化。在这里,我们在两种细胞培养模型中研究了线粒体DNA功能在神经干细胞早期分化过程中的重要性:一种是通过品系选择技术从胚胎干细胞中获得的CGR8-NS细胞系,另一种是从胚胎第14天的小鼠胎儿前脑中分离出的原代神经干细胞。我们检测到,NSC分化后,mtDNA含量急剧增加,使其mtDNA水平适应分化状态,但线粒体转录因子A的表达并没有随之发生变化。由于用溴化乙锭进行化学mtDNA耗竭无法从两种NSC类型中生成活的ρ°细胞系,我们用2'-3'-双脱氧胞苷抑制mtDNA聚合酶-γ来减少mtDNA复制,从而降低细胞中的mtDNA含量。在 NSC 分化过程中抑制 mtDNA 复制会减少神经发生,但不会减少神经胶质细胞的生成。在所使用的 NSC 模型中,mtDNA 的耗竭不会改变能量的产生/消耗或细胞活性氧(ROS)的含量。总之,mtDNA复制对胎儿NSCs的神经发生至关重要,但对神经胶质细胞的发生并不重要,其机制尚不清楚,但在很大程度上与能量/ROS代谢无关。
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引用次数: 0
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Development Growth & Differentiation
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