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Development of the brainstem respiratory circuit 脑干呼吸回路的发展
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-12-09 DOI: 10.1002/wdev.366
Meike E. van der Heijden, H. Zoghbi
The respiratory circuit is comprised of over a dozen functionally and anatomically segregated brainstem nuclei that work together to control respiratory rhythms. These respiratory rhythms emerge prenatally but only acquire vital importance at birth, which is the first time the respiratory circuit faces the sole responsibility for O2/CO2 homeostasis. Hence, the respiratory circuit has little room for trial‐and‐error‐dependent fine tuning and relies on a detailed genetic blueprint for development. This blueprint is provided by transcription factors that have specific spatiotemporal expression patterns along the rostral‐caudal or dorsal‐ventral axis of the developing brainstem, in proliferating precursor cells and postmitotic neurons. Studying these transcription factors in mice has provided key insights into the functional segregation of respiratory control and the vital importance of specific respiratory nuclei. Many studies converge on just two respiratory nuclei that each have rhythmogenic properties during the prenatal period: the preBötzinger complex (preBötC) and retrotrapezoid nucleus/parafacial nucleus (RTN/pF). Here, we discuss the transcriptional regulation that guides the development of these nuclei. We also summarize evidence showing that normal preBötC development is necessary for neonatal survival, and that neither the preBötC nor the RTN/pF alone is sufficient to sustain normal postnatal respiratory rhythms. Last, we highlight several studies that use intersectional genetics to assess the necessity of transcription factors only in subregions of their expression domain. These studies independently demonstrate that lack of RTN/pF neurons weakens the respiratory circuit, yet these neurons are not necessary for neonatal survival because developmentally related populations can compensate for abnormal RTN/pF function at birth.
呼吸回路由十几个功能上和解剖上分离的脑干核组成,它们共同控制呼吸节律。这些呼吸节律在胎儿时期就出现了,但只有在出生时才变得至关重要,这是呼吸回路第一次面对氧气/二氧化碳体内平衡的唯一责任。因此,呼吸回路几乎没有空间进行依赖于试验和错误的微调,而是依赖于详细的遗传蓝图进行发育。这一蓝图是由转录因子提供的,这些转录因子在发育中的脑干的吻侧-尾侧轴或背侧-腹侧轴、增殖的前体细胞和有丝分裂后神经元中具有特定的时空表达模式。在小鼠中研究这些转录因子为呼吸控制的功能分离和特定呼吸核的重要意义提供了关键的见解。许多研究都集中在两个呼吸核上,这两个核在产前都有节律性:preBötzinger复合体(preBötC)和后梯形核/面旁核(RTN/pF)。在这里,我们讨论指导这些细胞核发育的转录调控。我们还总结了正常的preBötC发育对新生儿生存是必要的证据,并且preBötC和RTN/pF单独都不足以维持正常的产后呼吸节律。最后,我们重点介绍了几项研究,这些研究使用交叉遗传学来评估转录因子仅在其表达域的亚区域中的必要性。这些独立的研究表明,RTN/pF神经元的缺乏削弱了呼吸回路,然而这些神经元对新生儿的生存并不是必需的,因为发育相关的群体可以在出生时补偿异常的RTN/pF功能。
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引用次数: 12
Stem cells in skeletal muscle growth and regeneration in amniotes and teleosts: Emerging themes 干细胞在羊膜动物和硬骨鱼骨骼肌生长和再生中的作用:新兴主题
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-11-19 DOI: 10.1002/wdev.365
A. Ruparelia, Dhanushika Ratnayake, P. Currie
Skeletal muscle is a contractile, postmitotic tissue that retains the capacity to grow and regenerate throughout life in amniotes and teleost. Both muscle growth and regeneration are regulated by obligate tissue resident muscle stem cells. Given that considerable knowledge exists on the myogenic process, recent studies have focused on examining the molecular markers of muscle stem cells, and on the intrinsic and extrinsic signals regulating their function. From this, two themes emerge: firstly, muscle stem cells display remarkable heterogeneity not only with regards to their gene expression profile, but also with respect to their behavior and function; and secondly, the stem cell niche is a critical regulator of muscle stem cell function during growth and regeneration. Here, we will address the current understanding of these emerging themes with emphasis on the distinct processes used by amniotes and teleost, and discuss the challenges and opportunities in the muscle growth and regeneration fields.
骨骼肌是一种可收缩的有丝分裂后组织,在羊膜动物和硬骨鱼的一生中保持生长和再生的能力。肌肉生长和再生都是由专性组织驻留肌肉干细胞调节的。鉴于对肌肉生成过程有相当多的了解,最近的研究主要集中在检查肌肉干细胞的分子标记,以及调节其功能的内在和外在信号。由此,出现了两个主题:首先,肌肉干细胞不仅在基因表达谱方面表现出显著的异质性,而且在行为和功能方面也表现出显著的异质性;其次,干细胞生态位是肌肉干细胞生长和再生过程中功能的关键调节器。在这里,我们将讨论当前对这些新兴主题的理解,重点是羊膜动物和硬骨鱼使用的不同过程,并讨论肌肉生长和再生领域的挑战和机遇。
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引用次数: 6
A critical look: Challenges in differentiating human pluripotent stem cells into desired cell types and organoids 批判性观察:将人类多能干细胞分化为所需细胞类型和类器官的挑战
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-11-19 DOI: 10.1002/wdev.368
J. Fowler, L. Ang, K. Loh
Too many choices can be problematic. This is certainly the case for human pluripotent stem cells (hPSCs): they harbor the potential to differentiate into hundreds of cell types; yet it is highly challenging to exclusively differentiate hPSCs into a single desired cell type. This review focuses on unresolved and fundamental questions regarding hPSC differentiation and critiquing the identity and purity of the resultant cell populations. These are timely issues in view of the fact that hPSC‐derived cell populations have or are being transplanted into patients in over 30 ongoing clinical trials. While many in vitro differentiation protocols purport to “mimic development,” the exact number and identity of intermediate steps that a pluripotent cell takes to differentiate into a given cell type in vivo remains largely unknown. Consequently, most differentiation efforts inevitably generate a heterogeneous cellular population, as revealed by single‐cell RNA‐sequencing and other analyses. The presence of unwanted cell types in differentiated hPSC populations does not portend well for transplantation therapies. This provides an impetus to precisely control differentiation to desired ends—for instance, by logically blocking the formation of unwanted cell types or by overexpressing lineage‐specifying transcription factors—or by harnessing technologies to selectively purify desired cell types. Conversely, approaches to differentiate three‐dimensional “organoids” from hPSCs intentionally generate heterogeneous cell populations. While this is intended to mimic the rich cellular diversity of developing tissues, whether all such organoids are spatially organized in a manner akin to native organs (and thus, whether they fully qualify as organoids) remains to be fully resolved.
选择太多可能会有问题。人类多能干细胞(hPSCs)当然是这样:它们具有分化为数百种细胞类型的潜力;然而将hPSC专门分化为单一所需的细胞类型是极具挑战性的。这篇综述的重点是关于hPSC分化的尚未解决的基本问题,以及对所得细胞群的身份和纯度的批评。鉴于hPSC衍生的细胞群已经或正在30多项正在进行的临床试验中移植到患者体内,这些都是及时的问题。尽管许多体外分化方案声称“模拟发育”,但多能干细胞在体内分化为特定细胞类型所需的中间步骤的确切数量和身份在很大程度上仍然未知。因此,正如单细胞RNA测序和其他分析所揭示的那样,大多数分化努力不可避免地会产生异质性细胞群体。分化的hPSC群体中存在不想要的细胞类型并不能很好地预示移植治疗。这为精确控制向所需目的的分化提供了动力,例如,通过逻辑上阻断不需要的细胞类型的形成,或通过过表达谱系特异性转录因子,或通过利用技术选择性纯化所需的细胞类型。相反,区分三维“类器官”和hPSC的方法有意产生异质性细胞群。虽然这是为了模拟发育中组织丰富的细胞多样性,但所有这些类器官是否都以类似于天然器官的方式在空间上组织(因此,它们是否完全符合类器官的资格)仍有待完全解决。
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引用次数: 29
Recent advancements in understanding fin regeneration in zebrafish 了解斑马鱼鳍再生的最新进展
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-11-14 DOI: 10.1002/wdev.367
I. Sehring, G. Weidinger
Zebrafish have the remarkable ability to fully regenerate a lost appendage, faithfully restoring its size, shape and tissue patterning. Studies over the past decades have identified mechanisms underlying the formation, spatial organization, and regenerative growth of the blastema, a pool of proliferative progenitor cells. The patterning of newly forming tissue is tightly regulated to ensure proper rebuilding of anatomy. Precise niche regulation of retinoic acid and sonic hedgehog signaling ensures adherence to ray—interray boundaries. The molecular underpinnings of systems underlying re‐establishment of pre‐amputation size and shape (positional information) are also slowly starting to emerge. Osteoblasts play an important role as a cellular source of regenerating skeletal elements, and in zebrafish both osteoblast dedifferentiation as well as de novo osteoblast formation occurs. Both dedifferentiation and proliferation are tightly controlled, which makes it interesting to compare it to tumorigenesis, and to identify potential players involved in these processes.
斑马鱼具有完全再生失去的附属物的非凡能力,忠实地恢复其大小,形状和组织图案。过去几十年的研究已经确定了囊胚形成、空间组织和再生生长的机制,囊胚是一个增殖祖细胞池。新形成组织的模式受到严格调控,以确保解剖结构的适当重建。维甲酸和超音刺猬信号的精确生态位调节确保了射线间边界的依从性。截肢前大小和形状(位置信息)重建系统的分子基础也慢慢开始出现。成骨细胞作为再生骨骼元素的细胞来源发挥着重要作用,在斑马鱼中,成骨细胞既发生去分化,也发生新生成骨细胞形成。去分化和增殖都受到严格控制,这使得将其与肿瘤发生进行比较并确定参与这些过程的潜在参与者变得有趣。
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引用次数: 61
Drug screening in Drosophila; why, when, and when not? 果蝇药物筛选;为什么,什么时候,什么时候不是?
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-11-01 Epub Date: 2019-05-05 DOI: 10.1002/wdev.346
Tin Tin Su

The best global seller among oncology drugs in 2018 is lenalidomide, an analog of thalidomide. It took 53 years and a circuitous route from the discovery of thalidomide to approval of an analog for use in treatment of cancer. We understand now a lot more about the genetic and molecular basis of diseases than we did in 1953 when thalidomide was discovered. We have also no shortage of chemical libraries with hundreds of thousands of compounds, both synthetic and natural. What we need are better ways to search among these rich resources for compounds with the potential to do what we want them to do. This review summarizes examples from the literature that make Drosophila melanogaster a good model to screen for drugs, and discusses knowledge gaps and technical challenges that make Drosophila models not as widely used as they could or should be. This article is categorized under: Technologies > Analysis of Cell, Tissue, and Animal Phenotypes.

2018年全球最畅销的肿瘤药物是沙利度胺的类似物来那度胺。从发现沙利度胺到批准一种类似药物用于治疗癌症,经历了53年的曲折历程。与1953年发现沙利度胺时相比,我们现在对疾病的遗传和分子基础有了更多的了解。我们也不缺化学文库,里面有成千上万的化合物,既有合成的,也有天然的。我们需要的是更好的方法,在这些丰富的资源中寻找具有我们想要的功能的化合物。这篇综述总结了文献中使果蝇成为药物筛选良好模型的例子,并讨论了使果蝇模型不能得到广泛应用的知识空白和技术挑战。本文分类如下:技术>细胞、组织和动物表型分析。
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引用次数: 26
Reflections on the use of protein binders to study protein function in developmental biology. 利用蛋白质结合物研究发育生物学中蛋白质功能的思考。
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-11-01 Epub Date: 2019-07-02 DOI: 10.1002/wdev.356
Gustavo Aguilar, M Alessandra Vigano, Markus Affolter, Shinya Matsuda

Studies in the field of developmental biology aim to unravel how a fertilized egg develops into an adult organism and how proteins and other macromolecules work together during this process. With regard to protein function, most of the developmental studies have used genetic and RNA interference approaches, combined with biochemical analyses, to reach this goal. However, there always remains much room for interpretation on how a given protein functions, because proteins work together with many other molecules in complex regulatory networks and it is not easy to reveal the function of one given protein without affecting the networks. Likewise, it has remained difficult to experimentally challenge and/or validate the proposed concepts derived from mutant analyses without tools that directly manipulate protein function in a predictable manner. Recently, synthetic tools based on protein binders such as scFvs, nanobodies, DARPins, and others have been applied in developmental biology to directly manipulate target proteins in a predicted manner. Although such tools would have a great impact in filling the gap of knowledge between mutant phenotypes and protein functions, careful investigations are required when applying functionalized protein binders to fundamental questions in developmental biology. In this review, we first summarize how protein binders have been used in the field, and then reflect on possible guidelines for applying such tools to study protein functions in developmental biology. This article is categorized under: Technologies > Analysis of Proteins Establishment of Spatial and Temporal Patterns > Gradients Invertebrate Organogenesis > Flies.

发育生物学领域的研究旨在揭示受精卵如何发育成成年生物体,以及蛋白质和其他大分子在此过程中如何共同作用。关于蛋白质的功能,大多数发育研究都采用遗传和RNA干扰方法,结合生化分析来达到这一目标。然而,对于一个给定的蛋白质是如何发挥作用的,仍然有很大的解释空间,因为蛋白质在复杂的调节网络中与许多其他分子一起工作,在不影响网络的情况下揭示一个给定蛋白质的功能是不容易的。同样,如果没有以可预测的方式直接操纵蛋白质功能的工具,在实验上挑战和/或验证从突变分析中得出的提出的概念仍然很困难。近年来,基于蛋白质结合物的合成工具如scFvs、纳米体、DARPins等已被应用于发育生物学中,以预测的方式直接操纵靶蛋白。尽管这些工具将在填补突变表型和蛋白质功能之间的知识空白方面产生重大影响,但在将功能化蛋白质结合物应用于发育生物学的基本问题时,需要进行仔细的研究。在这篇综述中,我们首先总结了蛋白质结合物在该领域的应用,然后反思了在发育生物学中应用这些工具研究蛋白质功能的可能指导方针。本文分类为:技术>蛋白质时空格局分析>无脊椎动物器官发生梯度>苍蝇。
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引用次数: 12
Methods for the study of innate immunity in Drosophila melanogaster. 黑腹果蝇先天免疫的研究方法。
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-09-01 Epub Date: 2019-04-16 DOI: 10.1002/wdev.344
Katia Troha, Nicolas Buchon

From flies to humans, many components of the innate immune system have been conserved during metazoan evolution. This foundational observation has allowed us to develop Drosophila melanogaster, the fruit fly, into a powerful model to study innate immunity in animals. Thanks to an ever-growing arsenal of genetic tools, an easily manipulated genome, and its winning disposition, Drosophila is now employed to study not only basic molecular mechanisms of pathogen recognition and immune signaling, but also the nature of physiological responses activated in the host by microbial challenge and how dysregulation of these processes contributes to disease. Here, we present a collection of methods and protocols to challenge the fly with an assortment of microbes, both systemically and orally, and assess its humoral, cellular, and epithelial response to infection. Our review covers techniques for measuring the reaction to microbial infection both qualitatively and quantitatively. Specifically, we describe survival, bacterial load, BLUD (a measure of disease tolerance), phagocytosis, melanization, clotting, and ROS production assays, as well as efficient protocols to collect hemolymph and measure immune gene expression. We also offer an updated catalog of online resources and a collection of popular reporter lines and mutants to facilitate research efforts. This article is categorized under: Technologies > Analysis of Cell, Tissue, and Animal Phenotypes.

从苍蝇到人类,先天免疫系统的许多成分在后生动物进化过程中被保存下来。这一基础观察使我们能够将果蝇发展成为研究动物先天免疫的强大模型。由于遗传工具的不断增加,易于操作的基因组,以及它的胜利倾向,果蝇现在不仅用于研究病原体识别和免疫信号的基本分子机制,而且还用于研究微生物挑战在宿主中激活的生理反应的本质以及这些过程的失调如何导致疾病。在这里,我们提出了一系列方法和方案,以系统和口服的方式用各种微生物挑战苍蝇,并评估其体液,细胞和上皮对感染的反应。我们的综述涵盖了对微生物感染反应的定性和定量测量技术。具体来说,我们描述了生存、细菌负荷、bld(一种疾病耐受性的测量方法)、吞噬、黑化、凝血和ROS产生测定,以及收集血淋巴和测量免疫基因表达的有效方案。我们还提供在线资源的更新目录和流行的记者线和突变的集合,以促进研究工作。本文分类如下:技术>细胞、组织和动物表型分析。
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引用次数: 51
The amphipod crustacean Parhyale hawaiensis: An emerging comparative model of arthropod development, evolution, and regeneration. 片足类甲壳纲夏威夷Parhyale:节肢动物发展、进化和再生的新兴比较模型。
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-09-01 Epub Date: 2019-06-11 DOI: 10.1002/wdev.355
Dennis A Sun, Nipam H Patel

Recent advances in genetic manipulation and genome sequencing have paved the way for a new generation of research organisms. The amphipod crustacean Parhyale hawaiensis is one such system. Parhyale are easy to rear and offer large broods of embryos amenable to injection, dissection, and live imaging. Foundational work has described Parhyale embryonic development, while advancements in genetic manipulation using CRISPR-Cas9 and other techniques, combined with genome and transcriptome sequencing, have enabled its use in studies of arthropod development, evolution, and regeneration. This study introduces Parhyale development and life history, a catalog of techniques and resources for Parhyale research, and two case studies illustrating its power as a comparative research system. This article is categorized under: Comparative Development and Evolution > Evolutionary Novelties Adult Stem Cells, Tissue Renewal, and Regeneration > Regeneration Comparative Development and Evolution > Model Systems Comparative Development and Evolution > Body Plan Evolution.

基因操作和基因组测序的最新进展为新一代研究生物铺平了道路。片脚甲壳类动物夏威夷Parhyale hawaiensis就是这样一个系统。Parhyale很容易培养,并提供大量的胚胎,适合注射,解剖和实时成像。基础工作描述了Parhyale胚胎发育,而使用CRISPR-Cas9和其他技术的遗传操作的进步,结合基因组和转录组测序,使其能够用于节肢动物发育、进化和再生的研究。本研究介绍了Parhyale的发展和生命历程,介绍了Parhyale研究的技术和资源目录,以及两个案例研究,说明了其作为比较研究系统的力量。本文分类如下:比较发育和进化>进化新颖性成体干细胞、组织更新和再生>再生比较发育和进化>模型系统比较发育和进化>身体计划进化。
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引用次数: 20
Development of the thalamus: From early patterning to regulation of cortical functions. 丘脑的发育:从早期模式到皮质功能的调节。
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-09-01 Epub Date: 2019-04-29 DOI: 10.1002/wdev.345
Yasushi Nakagawa

The thalamus is a brain structure of the vertebrate diencephalon that plays a central role in regulating diverse functions of the cerebral cortex. In traditional view of vertebrate neuroanatomy, the thalamus includes three regions, dorsal thalamus, ventral thalamus, and epithalamus. Recent molecular embryological studies have redefined the thalamus and the associated axial nomenclature of the diencephalon in the context of forebrain patterning. This new view has provided a useful conceptual framework for studies on molecular mechanisms of patterning, neurogenesis and fate specification in the thalamus as well as the guidance mechanisms for thalamocortical axons. Additionally, the availability of genetic tools in mice has led to important findings on how thalamic development is linked to the development of other brain regions, particularly the cerebral cortex. This article will give an overview of the organization of the embryonic thalamus and how progenitor cells in the thalamus generate neurons that are organized into discrete nuclei. I will then discuss how thalamic development is orchestrated with the development of the cerebral cortex and other brain regions. This article is categorized under: Nervous System Development > Vertebrates: Regional Development Nervous System Development > Vertebrates: General Principles.

丘脑是脊椎动物间脑的一个脑结构,在调节大脑皮层的多种功能方面起着核心作用。在传统的脊椎动物神经解剖学观点中,丘脑包括三个区域:背丘脑、腹侧丘脑和上皮层。最近的分子胚胎学研究在前脑模式的背景下重新定义了丘脑和间脑相关的轴向命名。这一新观点为研究丘脑的模式化、神经发生和命运规范的分子机制以及丘脑皮质轴突的引导机制提供了有益的概念框架。此外,小鼠遗传工具的可用性已经导致了丘脑发育如何与其他大脑区域,特别是大脑皮层的发育联系在一起的重要发现。本文将概述胚胎丘脑的组织,以及丘脑中的祖细胞如何产生组织成离散核的神经元。然后,我将讨论丘脑的发育是如何与大脑皮层和其他大脑区域的发育协调一致的。本文分类如下:神经系统发育>脊椎动物:区域发育神经系统发育>脊椎动物:一般原理。
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引用次数: 33
Regulation of insulin and adipokinetic hormone/glucagon production in flies 果蝇胰岛素和脂肪动力学激素/胰高血糖素产生的调控
Q1 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-08-04 DOI: 10.1002/wdev.360
Muhammad Ahmad, Li He, N. Perrimon
Metabolic homeostasis is under strict regulation of humoral factors across various taxa. In particular, insulin and glucagon, referred to in Drosophila as Drosophila insulin‐like peptides (DILPs) and adipokinetic hormone (AKH), respectively, are key hormones that regulate metabolism in most metazoa. While much is known about the regulation of DILPs, the mechanisms regulating AKH/glucagon production is still poorly understood. In this review, we describe the various factors that regulate the production of DILPs and AKH and emphasize the need for future studies to decipher how energy homeostasis is governed in Drosophila.
代谢稳态是在各种类群的体液因子的严格调控下进行的。特别是胰岛素和胰高血糖素,在果蝇中分别被称为果蝇胰岛素样肽(DILPs)和脂肪动力学激素(AKH),是调节大多数后生动物代谢的关键激素。虽然我们对dilp的调节了解很多,但对AKH/胰高血糖素产生的调节机制仍然知之甚少。在这篇综述中,我们描述了调节dilp和AKH产生的各种因素,并强调需要未来的研究来解释果蝇是如何控制能量稳态的。
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引用次数: 40
期刊
Wiley Interdisciplinary Reviews: Developmental Biology
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