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Semi-automated, high-content imaging of drug transporter knockout sea urchin (Lytechinus pictus) embryos 药物转运体基因敲除海胆(Lytechinus pictus)胚胎的半自动高含量成像
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-12-12 DOI: 10.1002/jez.b.23231
Evan Tjeerdema, Yoon Lee, Rachel Metry, Amro Hamdoun

A defining feature of sea urchins is their extreme fecundity. Urchins produce millions of transparent, synchronously developing embryos, ideal for spatial and temporal analysis of development. This biological feature has been effectively utilized for ensemble measurement of biochemical changes. However, it has been underutilized in imaging studies, where single embryo measurements are used. Here we present an example of how stable genetics and high content imaging, along with machine learning-based image analysis, can be used to exploit the fecundity and synchrony of sea urchins in imaging-based drug screens. Building upon our recently created sea urchin ABCB1 knockout line, we developed a high-throughput assay to probe the role of this drug transporter in embryos. We used high content imaging to compare accumulation and toxicity of canonical substrates and inhibitors of the transporter, including fluorescent molecules and antimitotic cancer drugs, in homozygous knockout and wildtype embryos. To measure responses from the resulting image data, we used a nested convolutional neural network, which rapidly classified embryos according to fluorescence or cell division. This approach identified sea urchin embryos with 99.8% accuracy and determined two-cell and aberrant embryos with 96.3% and 89.1% accuracy, respectively. The results revealed that ABCB1 knockout embryos accumulated the transporter substrate calcein 3.09 times faster than wildtypes. Similarly, knockouts were 4.71 and 3.07 times more sensitive to the mitotic poisons vinblastine and taxol. This study paves the way for large scale pharmacological screens in the sea urchin embryo.

海胆的一个显著特征是它们的繁殖力极强。海胆产生数百万个透明的、同步发育的胚胎,非常适合进行时空发育分析。这一生物学特性已被有效地用于生化变化的集合测量。然而,它在成像研究中未得到充分利用,其中使用单胚胎测量。在这里,我们提出了一个例子,说明如何使用稳定的遗传学和高含量成像,以及基于机器学习的图像分析,在基于成像的药物筛选中利用海胆的繁殖能力和同步性。在我们最近创建的海胆ABCB1基因敲除系的基础上,我们开发了一种高通量测定方法来探测这种药物转运体在胚胎中的作用。我们使用高含量成像来比较纯合子敲除和野生型胚胎中典型底物和转运体抑制剂的积累和毒性,包括荧光分子和抗有丝分裂癌症药物。为了测量由此产生的图像数据的反应,我们使用了一个嵌套的卷积神经网络,该网络根据荧光或细胞分裂快速分类胚胎。该方法鉴定海胆胚胎的准确率为99.8%,鉴定双细胞和异常胚胎的准确率分别为96.3%和89.1%。结果表明,ABCB1基因敲除胚胎转运蛋白底物钙黄蛋白积累速度比野生型快3.09倍。同样,基因敲除对有丝分裂毒素长春花碱和紫杉醇的敏感性分别是前者的4.71倍和3.07倍。本研究为海胆胚胎的大规模药理筛选铺平了道路。
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引用次数: 0
Annelids as models of germ cell and gonad regeneration 作为生殖细胞和性腺再生模型的无脊椎动物
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-12-11 DOI: 10.1002/jez.b.23233
B. Duygu Özpolat

Germ cells (reproductive cells and their progenitors) give rise to the next generation in sexually reproducing organisms. The loss or removal of germ cells often leads to sterility in established research organisms such as the fruit fly, nematodes, frog, and mouse. The failure to regenerate germ cells in these organisms reinforced the dogma of germline–soma barrier in which germ cells are set-aside during embryogenesis and cannot be replaced by somatic cells. However, in stark contrast, many animals including segmented worms (annelids), hydrozoans, planaria, sea stars, sea urchins, and tunicates can regenerate germ cells. Here I review germ cell and gonad regeneration in annelids, a rich history of research that dates back to the early 20th century in this highly regenerative group. Examples include annelids from across the annelid phylogeny, across developmental stages, and reproductive strategies. Adult annelids regenerate germ cells as a part of regeneration, grafting, and asexual reproduction. Annelids can also recover germ cells after ablation of germ cell progenitors in the embryos. I present a framework to investigate cellular sources of germ cell regeneration in annelids, and discuss the literature that supports different possibilities within this framework, where germ–soma separation may or may not be preserved. With contemporary genetic-lineage tracing and bioinformatics tools, and several genetically enabled annelid models, we are at the brink of answering the big questions that puzzled many for over more than a century.

生殖细胞(生殖细胞及其祖细胞)是有性生殖生物的下一代。在果蝇、线虫、青蛙和小鼠等成熟的研究生物中,生殖细胞的丧失或移除往往会导致不育。这些生物的生殖细胞无法再生,这强化了生殖细胞-体细胞屏障的教条,即生殖细胞在胚胎发生过程中被搁置,不能被体细胞取代。然而,与此形成鲜明对比的是,许多动物,包括节肢动物(无脊椎动物)、水螅、扁形动物、海星、海胆和鳞鳃纲动物,都能再生生殖细胞。在这里,我将回顾环口纲动物的生殖细胞和性腺再生,这一研究历史悠久,可以追溯到 20 世纪初。例子包括环口纲动物的整个系统发育、各个发育阶段和繁殖策略。作为再生、嫁接和无性繁殖的一部分,成年环带动物可再生生殖细胞。无环动物还能在胚胎中的生殖细胞祖细胞被消减后恢复生殖细胞。我提出了一个研究环带动物生殖细胞再生细胞来源的框架,并讨论了支持该框架内不同可能性的文献,其中生殖细胞-体细胞分离可能会也可能不会被保留。有了当代的基因谱系追踪和生物信息学工具,以及几种通过基因改造的环带动物模型,我们即将回答一个多世纪以来困惑许多人的重大问题。
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引用次数: 0
Natural male hybrid common shrews with a very long chromosomal multivalent at meiosis appear not to be completely sterile 在减数分裂时具有很长多价染色体的天然雄性杂交鼩鼱似乎不是完全不育的。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-12-07 DOI: 10.1002/jez.b.23232
Sergey N. Matveevsky, Oxana L. Kolomiets, Nikolay A. Shchipanov, Svetlana V. Pavlova

Among 36 known chromosomal hybrid zones of the common shrew Sorex araneus, the Moscow–Seliger hybrid zone is of special interest because inter-racial complex heterozygotes (F1 hybrids) produce the longest meiotic configuration, consisting of 11 chromosomes with monobrachial homology (undecavalent or chain-of-eleven: CXI). Different studies suggest that such a multivalent may negatively affect meiotic progression and in general should significantly reduce fertility of hybrids. In this work, by immunocytochemical and electron microscopy methods, we investigated for the first time chromosome synapsis, recombination and meiotic silencing in pachytene spermatocytes of natural inter-racial heterozygous shrew males carrying CXI configurations. Despite some abnormalities detected in spermatocytes, such as associations of chromosomes, stretched centromeres, and the absence of recombination nodules in some arms of the multivalent, a large number of morphologically normal spermatozoa were observed. Possible low stringency of pachytene checkpoints may mean that even very long meiotic configurations do not cause complete sterility of such complex inter-racial heterozygotes.

在已知的36个普通鼩染色体杂交区中,莫斯科-塞利格杂交区特别有趣,因为种族间复杂杂合子(F1杂交)产生最长的减数分裂构型,由11条染色体组成,具有单臂同源性(十一链:CXI)。不同的研究表明,这样的多价可能会对减数分裂进程产生负面影响,并且通常会显著降低杂交种的育性。本文采用免疫细胞化学和电镜技术,首次研究了携带CXI构型的自然种间杂合雄性鼩猴粗线精母细胞的染色体突触、重组和减数分裂沉默。尽管在精母细胞中检测到一些异常,如染色体关联,着丝粒拉伸,以及在多价染色体的一些臂中没有重组结节,但仍观察到大量形态正常的精子。粗线蛋白检查点可能的低严格性可能意味着即使很长的减数分裂构型也不会导致这种复杂的种族间杂合子的完全不育。
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引用次数: 0
Transitioning from a research protocol to a scalable applied pathway for Xenopus laevis sperm cryopreservation at a national stock center: The effect of cryoprotectants 从研究方案到国家储备中心非洲爪蟾精子冷冻保存的可扩展应用途径的过渡:冷冻保护剂的作用。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-11-20 DOI: 10.1002/jez.b.23228
Lucía Arregui, Jack C. Koch, Terrence R. Tiersch

Sperm cryopreservation is a critical tool for safeguarding and managing valuable genetic resources. Protocols for cryopreservation of Xenopus laevis sperm were available but lacking sperm quality evaluation and scalability and the outcomes were inconsistent. The goal of this study was to begin developing a center-level cryopreservation pathway for this species by integrating French straws as containers that would facilitate germplasm repository development. The objectives were to analyze the effect of: (1) three sperm concentrations (33, 50, and 100 × 106 sperm/mL) on post-thaw fertilization, (2) three final concentrations (2.5%, 5%, and 10%) of dimethyl sulfoxide, methanol, and dimethylformamide (DMFA) on sperm membrane integrity of fresh and frozen samples, (3) two concentrations (5% and 10%) of DMFA with and without 5% sucrose at four cooling rates (5, 10, 20, and 40°C/min) on sperm membrane integrity and motility, and (4) egg exposure to different concentrations of DMFA on fertilization. Few differences in sperm viability were found among fresh samples incubated in cryoprotectants, but thawed samples frozen in methanol or DMFA presented higher membrane integrity. Samples frozen in 10% DMFA at 20°C/min showed higher membrane integrity (60 ± 7%) than other DMFA concentrations and cooling rates, and the same total motility (30 ± 7%) as at 10°C/min. Higher DMFA concentrations (10%–13%) were detrimental for embryo development compared to lower concentrations (<6%). This study provided a reliable protocol for sperm cryopreservation in Xenopus laevis to yield an application pathway with potential for high throughput that can be used as a roadmap for work with other species.

精子冷冻保存是保护和管理宝贵遗传资源的重要手段。目前已有非洲爪蟾精子冷冻保存方案,但缺乏精子质量评价和可扩展性,结果不一致。本研究的目的是通过将法国吸管作为容器,促进种质资源库的开发,开始开发该物种的中心级低温保存途径。目的是分析以下措施的效果:(1)三个精子浓度(33、50和100×106精子/毫升)post-thaw受精,(2)三个最终浓度(2.5%,5%,和10%)二甲亚砜、甲醇、二甲基甲酰胺(DMFA)精子膜完整性的新鲜和冷冻样品,(3)两种DMFA浓度(5%和10%),没有5%蔗糖在四个冷却率(5、10、20和40°C / min)对精子膜完整性和能动性,和(4)蛋暴露于不同浓度的DMFA受精。在冷冻保护剂中孵育的新鲜样品中发现精子活力差异不大,但在甲醇或DMFA中冷冻的解冻样品具有更高的膜完整性。以20°C/min冷冻10% DMFA的样品,其膜完整性(60±7%)高于其他DMFA浓度和冷却速率,总运动性(30±7%)与10°C/min相同。较高的DMFA浓度(10%-13%)对胚胎发育不利,较低的DMFA浓度(
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引用次数: 0
Henri de Lacaze-Duthiers and the ascidian hypothesis 亨利-德-拉卡泽-杜蒂耶和腹水假说。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-11-16 DOI: 10.1002/jez.b.23226
Catherine Jessus, Vincent Laudet

In 1830, Cuvier and Geoffroy Saint-Hilaire confronted each other in a famous debate on the unity of the animal kingdom, which permeated the zoology of the 19th century. From that time, a growing number of naturalists attempted to understand the large-scale relationships among animals. And among all the questions, that of the origin of vertebrates was one of the most controversial. Analytical methods based on comparative anatomy, embryology and paleontology were developed to identify convincing homologies that would reveal a logical sequence of events for the evolution of an invertebrate into the first vertebrate. Within this context, several theories have clashed on the question of the identity of the ancestor of vertebrates. Among the proposals, a group of rather discrete organisms, the ascidians, played a central role. Because he had discovered an ascidian with a particularly atypical larval development, the Molgula, Henri de Lacaze-Duthiers, a rigorous and meticulous naturalist, became involved in the ascidian hypothesis. While the visionary mind of Lacaze-Duthiers led him to establish a particularly innovative methodology and the first marine biology station in Europe, at Roscoff, the tailless tadpole of the Molgula prevented him from recognizing the ancestor of vertebrates. This old 19th century story echoes the ever-present questions driving the field of Eco-Evo-Devo.

1830年,居维叶和若弗鲁瓦·圣伊莱尔就动物王国的统一性进行了一场著名的辩论,这场辩论渗透到19世纪的动物学中。从那时起,越来越多的博物学家试图了解动物之间的大规模关系。在所有的问题中,脊椎动物的起源是最有争议的问题之一。基于比较解剖学、胚胎学和古生物学的分析方法被开发出来,以确定令人信服的同源性,从而揭示无脊椎动物向第一个脊椎动物进化的逻辑序列。在这种背景下,几种理论在脊椎动物祖先的身份问题上发生了冲突。在这些建议中,一组相当分散的生物,海鞘,发挥了核心作用。由于亨利·德·拉卡兹-杜蒂耶斯发现了一种具有特别非典型幼虫发育的海鞘,即Molgula,这位严谨细致的博物学家开始参与海鞘假说。虽然拉卡兹-杜蒂耶斯的远见卓识使他在罗斯科夫建立了一种特别创新的方法,并在欧洲建立了第一个海洋生物学站,但Molgula的无尾蝌蚪使他无法识别脊椎动物的祖先。这个古老的19世纪的故事呼应了驱动生态-生态-发展领域的始终存在的问题。
{"title":"Henri de Lacaze-Duthiers and the ascidian hypothesis","authors":"Catherine Jessus,&nbsp;Vincent Laudet","doi":"10.1002/jez.b.23226","DOIUrl":"10.1002/jez.b.23226","url":null,"abstract":"<p>In 1830, Cuvier and Geoffroy Saint-Hilaire confronted each other in a famous debate on the unity of the animal kingdom, which permeated the zoology of the 19th century. From that time, a growing number of naturalists attempted to understand the large-scale relationships among animals. And among all the questions, that of the origin of vertebrates was one of the most controversial. Analytical methods based on comparative anatomy, embryology and paleontology were developed to identify convincing homologies that would reveal a logical sequence of events for the evolution of an invertebrate into the first vertebrate. Within this context, several theories have clashed on the question of the identity of the ancestor of vertebrates. Among the proposals, a group of rather discrete organisms, the ascidians, played a central role. Because he had discovered an ascidian with a particularly atypical larval development, the <i>Molgula</i>, Henri de Lacaze-Duthiers, a rigorous and meticulous naturalist, became involved in the ascidian hypothesis. While the visionary mind of Lacaze-Duthiers led him to establish a particularly innovative methodology and the first marine biology station in Europe, at Roscoff, the tailless tadpole of the <i>Molgula</i> prevented him from recognizing the ancestor of vertebrates. This old 19th century story echoes the ever-present questions driving the field of Eco-Evo-Devo.</p>","PeriodicalId":15682,"journal":{"name":"Journal of experimental zoology. Part B, Molecular and developmental evolution","volume":"342 1","pages":"7-20"},"PeriodicalIF":2.2,"publicationDate":"2023-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jez.b.23226","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136397687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Functional analyses of the polycomb-group genes in sea lamprey embryos undergoing programmed DNA loss 经历程序性DNA丢失的七叶树胚胎中多梳群基因的功能分析。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-10-30 DOI: 10.1002/jez.b.23225
Cody Saraceno, Vladimir A. Timoshevskiy, Jeramiah J. Smith

During early development, sea lamprey embryos undergo programmatic elimination of DNA from somatic progenitor cells in a process termed programmed genome rearrangement (PGR). Eliminated DNA eventually becomes condensed into micronuclei, which are then physically degraded and permanently lost from the cell. Previous studies indicated that many of the genes eliminated during PGR have mammalian homologs that are bound by polycomb repressive complex (PRC) in embryonic stem cells. To test whether PRC components play a role in the faithful elimination of germline-specific sequences, we used a combination of CRISPR/Cas9 and lightsheet microscopy to investigate the impact of gene knockouts on early development and the progression through stages of DNA elimination. Analysis of knockout embryos for the core PRC2 subunits EZH, SUZ12, and EED show that disruption of all three genes results in an increase in micronucleus number, altered distribution of micronuclei within embryos, and an increase in micronucleus volume in mutant embryos. While the upstream events of DNA elimination are not strongly impacted by loss of PRC2 components, this study suggests that PRC2 plays a role in the later stages of elimination related to micronucleus condensation and degradation. These findings also suggest that other genes/epigenetic pathways may work in parallel during DNA elimination to mediate chromatin structure, accessibility, and the ultimate loss of germline-specific DNA.

在早期发育过程中,七叶树胚胎经历了从体细胞祖细胞中程序性清除DNA的过程,这一过程被称为程序性基因组重排(PGR)。被清除的DNA最终凝结成微核,然后被物理降解并从细胞中永久丢失。先前的研究表明,在PGR过程中消除的许多基因具有哺乳动物同源物,这些同源物通过胚胎干细胞中的多梳抑制复合物(PRC)结合。为了测试PRC成分是否在种系特异性序列的忠实消除中发挥作用,我们使用CRISPR/Cas9和光片显微镜的组合来研究基因敲除对早期发育和DNA消除阶段进展的影响。对敲除胚胎的核心PRC2亚基EZH、SUZ12和EED的分析表明,所有三个基因的破坏都会导致微核数量增加,微核在胚胎内的分布改变,突变胚胎的微核体积增加。虽然DNA消除的上游事件不会受到PRC2成分损失的强烈影响,但本研究表明,PRC2在与微核浓缩和降解相关的消除后期发挥作用。这些发现还表明,在DNA消除过程中,其他基因/表观遗传途径可能并行工作,以介导染色质结构、可及性和种系特异性DNA的最终损失。
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引用次数: 0
Transcriptional background effects on a tumor driver gene in different pigment cell types of medaka 不同色素细胞类型对肿瘤驱动基因的转录背景效应。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-10-25 DOI: 10.1002/jez.b.23224
Shahad Abdulsahib, William Boswell, Mikki Boswell, Markita Savage, Manfred Schartl, Yuan Lu

The Xiphophorus melanoma receptor kinase gene, xmrk, is a bona fide oncogene driving melanocyte tumorigenesis of Xiphophorus fish. When ectopically expressed in medaka, it not only induces development of several pigment cell tumor types in different strains of medaka but also induces different tumor types within the same animal, suggesting its oncogenic activity has a transcriptomic background effect. Although the central pathways that xmrk utilizes to lead to melanomagenesis are well documented, genes and genetic pathways that modulate the oncogenic effect and alter the course of disease have not been studied so far. To understand how the genetic networks between different histocytes of xmrk-driven tumors are composed, we isolated two types of tumors, melanoma and xanthoerythrophoroma, from the same xmrk transgenic medaka individuals, established the transcriptional profiles of both xmrk-driven tumors, and compared (1) genes that are co-expressed with xmrk in both tumor types, and (2) differentially expressed genes and their associated molecular functions, between the two tumor types. Transcriptomic comparisons between the two tumor types show melanoma and xanthoerythrophoroma are characterized by transcriptional features representing varied functions, indicating distinct molecular interactions between the driving oncogene and the cell-type-specific transcriptomes. Melanoma tumors exhibit gene signatures that are relevant to proliferation and invasion, while xanthoerythrophoroma tumors are characterized by expression profiles related to metabolism and DNA repair. We conclude the transcriptomic backgrounds, exemplified by cell-type-specific genes that are downstream of xmrk effected signaling pathways, contribute the potential to change the course of tumor development and may affect overall tumor outcomes.

剑鱼黑色素瘤受体激酶基因xmrk是驱动剑鱼黑素细胞肿瘤发生的真正致癌基因。当它在青金石中异位表达时,它不仅在不同的青金石菌株中诱导几种色素细胞肿瘤类型的发展,而且在同一动物中诱导不同的肿瘤类型,这表明它的致癌活性具有转录组背景效应。尽管xmrk用来导致黑色素瘤的中心途径已经有了很好的记录,但到目前为止,调节致癌作用和改变疾病进程的基因和遗传途径还没有得到研究。为了了解xmrk驱动的肿瘤的不同组织细胞之间的遗传网络是如何组成的,我们从同一个xmrk转基因梅达卡个体中分离出两种类型的肿瘤,黑色素瘤和黄红细胞瘤,建立了两种xmrk驱动肿瘤的转录谱,并比较了(1)在两种肿瘤类型中与xmrk共表达的基因,以及(2)两种肿瘤类型之间的差异表达基因及其相关分子功能。两种肿瘤类型之间的转录组学比较表明,黑色素瘤和黄红细胞瘤的特征是代表不同功能的转录特征,表明驱动癌基因和细胞类型特异性转录组之间存在不同的分子相互作用。黑色素瘤肿瘤表现出与增殖和侵袭相关的基因特征,而黄红细胞瘤肿瘤的特征是与代谢和DNA修复相关的表达谱。我们得出结论,转录组背景,以xmrk影响的信号通路下游的细胞类型特异性基因为例,有可能改变肿瘤发展过程,并可能影响整体肿瘤结果。
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引用次数: 0
Suppressors of cGAS-STING are downregulated during fin-limb regeneration and aging in aquatic vertebrates cGAS-STING抑制因子在水生脊椎动物的鳍肢再生和衰老过程中下调。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-10-25 DOI: 10.1002/jez.b.23227
Sabateeshan Mathavarajah, Andrew W. Thompson, Matthew R. Stoyek, T. Alexander Quinn, Stéphane Roy, Ingo Braasch, Graham Dellaire

During the early stages of limb and fin regeneration in aquatic vertebrates (i.e., fishes and amphibians), blastema undergo transcriptional rewiring of innate immune signaling pathways to promote immune cell recruitment. In mammals, a fundamental component of innate immune signaling is the cytosolic DNA sensing pathway, cGAS-STING. However, to what extent the cGAS-STING pathway influences regeneration in aquatic anamniotes is unknown. In jawed vertebrates, negative regulation of cGAS-STING activity is accomplished by suppressors of cytosolic DNA such as Trex1, Pml, and PML-like exon 9 (Plex9) exonucleases. Here, we examine the expression of these suppressors of cGAS-STING, as well as inflammatory genes and cGAS activity during caudal fin and limb regeneration using the spotted gar (Lepisosteus oculatus) and axolotl (Ambystoma mexicanum) model species, and during age-related senescence in zebrafish (Danio rerio). In the regenerative blastema of wounded gar and axolotl, we observe increased inflammatory gene expression, including interferon genes and interleukins 6 and 8. We also observed a decrease in axolotl Trex1 and gar pml expression during the early phases of wound healing which correlates with a dramatic increase in cGAS activity. In contrast, the plex9.1 gene does not change in expression during wound healing in gar. However, we observed decreased expression of plex9.1 in the senescing cardiac tissue of aged zebrafish, where 2′3′-cGAMP levels are elevated. Finally, we demonstrate a similar pattern of Trex1, pml, and plex9.1 gene regulation across species in response to exogenous 2′3′-cGAMP. Thus, during the early stages of limb-fin regeneration, Pml, Trex1, and Plex9.1 exonucleases are downregulated, presumably to allow an evolutionarily ancient cGAS-STING activity to promote inflammation and the recruitment of immune cells.

在水生脊椎动物(即鱼类和两栖动物)四肢和鳍再生的早期阶段,芽基细胞经历先天免疫信号通路的转录重组,以促进免疫细胞的募集。在哺乳动物中,先天免疫信号传导的一个基本组成部分是胞质DNA传感通路cGAS-STING。然而,cGAS-STING途径在多大程度上影响水生蝾螈的再生尚不清楚。在有颌脊椎动物中,cGAS-STING活性的负调控是通过胞质DNA的抑制剂如Trex1、Pml和Pml样外显子9(Plex9)外切酶来实现的。在这里,我们使用斑点gar(Lepisosteus oculatus)和axolotl(Ambystoma mexicanum)模型物种,以及斑马鱼(Danio rerio)年龄相关衰老过程中,检测了cGAS STING的这些抑制剂的表达,以及炎症基因和cGAS活性。在受伤的gar和axolotl的再生芽细胞中,我们观察到炎症基因表达增加,包括干扰素基因和白细胞介素6和8。我们还观察到,在伤口愈合的早期阶段,axolotl Trex1和gar pml的表达减少,这与cGAS活性的显著增加有关。相反,在gar的伤口愈合过程中,plex9.1基因的表达没有改变。然而,我们观察到老年斑马鱼衰老心脏组织中plex9.1的表达减少,其中2'3'-cGAMP水平升高。最后,我们证明了Trex1、pml和plex9.1基因在物种间对外源性2'3'-cGAMP的调节模式相似。因此,在肢鳍再生的早期阶段,Pml、Trex1和Plex9.1外切酶被下调,可能是为了使进化上古老的cGAS STING活性能够促进炎症和免疫细胞的募集。
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引用次数: 0
Using Xenopus to discover new candidate genes involved in BOR and other congenital hearing loss syndromes 利用非洲爪蟾发现与BOR和其他先天性听力损失综合征有关的新的候选基因。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-10-13 DOI: 10.1002/jez.b.23222
Scott J. Neal, Anindita Rajasekaran, Nisveta Jusić, Louis Taylor, Mai Read, Dominique Alfandari, Francesca Pignoni, Sally A. Moody

Hearing in infants is essential for brain development, acquisition of verbal language skills, and development of social interactions. Therefore, it is important to diagnose hearing loss soon after birth so that interventions can be provided as early as possible. Most newborns in the United States are screened for hearing deficits and commercially available next-generation sequencing hearing loss panels often can identify the causative gene, which may also identify congenital defects in other organs. One of the most prevalent autosomal dominant congenital hearing loss syndromes is branchio-oto-renal syndrome (BOR), which also presents with defects in craniofacial structures and the kidney. Currently, mutations in three genes, SIX1, SIX5, and EYA1, are known to be causative in about half of the BOR patients that have been tested. To uncover new candidate genes that could be added to congenital hearing loss genetic screens, we have combined the power of Drosophila mutants and protein biochemical assays with the embryological advantages of Xenopus, a key aquatic animal model with a high level of genomic similarity to human, to identify potential Six1 transcriptional targets and interacting proteins that play a role during otic development. We review our transcriptomic, yeast 2-hybrid, and proteomic approaches that have revealed a large number of new candidates. We also discuss how we have begun to identify how Six1 and co-factors interact to direct developmental events necessary for normal otic development.

婴儿的听力对大脑发育、语言技能的习得和社交互动的发展至关重要。因此,重要的是在出生后立即诊断听力损失,以便尽早提供干预措施。在美国,大多数新生儿都会接受听力缺陷筛查,商业上可买到的下一代测序听力损失小组通常可以识别致病基因,这也可能识别其他器官的先天性缺陷。最常见的常染色体显性遗传先天性听力损失综合征之一是鳃-肾综合征(BOR),它也表现为颅面结构和肾脏的缺陷。目前,已知在大约一半接受测试的BOR患者中,SIX1、SIX5和EYA1三个基因的突变是致病原因。为了发现可以添加到先天性听力损失基因筛选中的新的候选基因,我们将果蝇突变体和蛋白质生化分析的能力与非洲爪蟾的胚胎学优势相结合,非洲爪蟾是一种与人类基因组高度相似的关键水生动物模型,以鉴定在耳发育过程中发挥作用的潜在Six1转录靶标和相互作用蛋白。我们回顾了我们的转录组学、酵母2-杂交和蛋白质组学方法,这些方法揭示了大量新的候选者。我们还讨论了我们是如何开始确定Six1和共因子如何相互作用来指导正常耳发育所需的发育事件的。
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引用次数: 0
The paradise fish, an advanced animal model for behavioral genetics and evolutionary developmental biology 天堂鱼,行为遗传学和进化发育生物学的高级动物模型。
IF 2.2 3区 生物学 Q3 DEVELOPMENTAL BIOLOGY Pub Date : 2023-10-11 DOI: 10.1002/jez.b.23223
Nóra Szabó, Erika Fodor, Zoltán Varga, Anita Tarján-Rácz, Kata Szabó, Ádám Miklósi, Máté Varga

Paradise fish (Macropodus opercularis) is an air-breathing freshwater fish species with a signature labyrinth organ capable of extracting oxygen from the air that helps these fish to survive in hypoxic environments. The appearance of this evolutionary innovation in anabantoids resulted in a rewired circulatory system, but also in the emergence of species-specific behaviors, such as territorial display, courtship and parental care in the case of the paradise fish. Early zoologists were intrigued by the structure and function of the labyrinth apparatus and a series of detailed descriptive histological studies at the beginning of the 20th century revealed the ontogenesis and function of this specialized system. A few decades later, these fish became the subject of numerous ethological studies, and detailed ethograms of their behavior were constructed. These latter studies also demonstrated a strong genetic component underlying their behavior, but due to lack of adequate molecular tools, the fine genetic dissection of the behavior was not possible at the time. The technological breakthroughs that transformed developmental biology and behavioral genetics in the past decades, however, give us now a unique opportunity to revisit these old questions. Building on the classic descriptive studies, the new methodologies will allow us to follow the development of the labyrinth apparatus at a cellular resolution, reveal the genes involved in this process and also the genetic architecture behind the complex behaviors that we can observe in this species.

天堂鱼(Macropodus opercularis)是一种呼吸空气的淡水鱼类,具有标志性的迷宫器官,能够从空气中提取氧气,帮助这些鱼类在缺氧环境中生存。这一进化创新在类动物中的出现导致了循环系统的重新连接,但也导致了物种特异性行为的出现,如天堂鱼的领地展示、求偶和父母照顾。早期动物学家对迷宫器的结构和功能很感兴趣,20世纪初的一系列详细的描述性组织学研究揭示了这个特殊系统的个体发生和功能。几十年后,这些鱼成为了许多行为学研究的对象,并构建了它们行为的详细行为图谱。后一项研究也证明了他们行为背后有强大的遗传成分,但由于缺乏足够的分子工具,当时还不可能对这种行为进行精细的遗传解剖。然而,在过去几十年里,改变了发育生物学和行为遗传学的技术突破,现在给了我们一个重新审视这些古老问题的独特机会。在经典描述性研究的基础上,新方法将使我们能够以细胞分辨率跟踪迷宫装置的发展,揭示参与这一过程的基因,以及我们可以在该物种中观察到的复杂行为背后的遗传结构。
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Journal of experimental zoology. Part B, Molecular and developmental evolution
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