Novel zebrafish approaches to investigate leaderless protein secretion in vivo

研究斑马鱼体内无领导蛋白分泌的新方法

• 批准号: BB/L000830/1
• 负责人: Heather Wilson
• 金额: $ 47.57万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL000830%2F1
• 关键词: Novel; zebrafish; approaches; investigate; leaderless

All cells within the body traffic their proteins either to compartments within the cell, or to the cell surface. At the cell surface they can either remain attached to the cell or they are secreted so that these proteins can be "seen" outside the cell, and away from the cell. The vast majority of surface or secreted proteins have a specific signal directing them to automatically traffic there. Recently it has been found that a small number of proteins, which play a role outside the cell but were assumed to be externalised only upon cell death, are in fact secreted under certain circumstances. One group of secreted proteins are inflammatory cytokines (messengers), critical in signaling to blood cells the location of an injury or infection and so essential to maintain a healthy immune system throughout life. We now know that activation of the damage-sensing "P2X7" cell surface receptor causes release (secretion) of inflammatory mediators (specifically "interleukin-1) into the bloodstream. Interestingly, when interleukin-1 exits the cell, it is enveloped in small, protected membrane packets, that we have termed "microvesicles". These packets are likely to prevent this highly inflammatory cytokine from acting at random sites within the body, and may provide a means of targeting it to particular cells where the microvesicles can dock. The cellular machinery coordinating microvesicle formation and cytokine secretion is complex. Several pathways involved in this process have been proposed, based on studies using a single type of cell grown in culture. We have developed a new system where we can study fluorescent cytokine in a whole live organism, namely the transparent zebrafish larva. For the first time, this offers us the opportunity to view the cells that secrete interleukin-1, and to see where interleukin-1 is targeted in a live organism. In this project we will optimise computational approaches to quantify vesicle formation, tracking and targeting in a whole organism using our fluorescent zebrafish lines. We will then test which cellular components are responsible for interleukin-1 secretion by adding inhibitors to these components and measuring changes in response to an infection. We will also use these fluorescent zebrafish to screen for small molecules that are effective in blocking inflammation, so that we can identify the critical parts of the cell that are required to for a healthy immune response. In the future, beyond this project, this will help us to develop new therapies to fight disease.

体内的所有细胞都将它们的蛋白质运输到细胞内的隔间或细胞表面。在细胞表面，它们既可以附着在细胞上，也可以被分泌出来，这样这些蛋白质就可以在细胞外和远离细胞的地方被“看到”。绝大多数表面或分泌的蛋白质都有一个特定的信号，引导它们自动进入那里。最近发现，在细胞外发挥作用但被认为只有在细胞死亡时才外化的少量蛋白质，实际上在某些情况下是分泌的。一组分泌的蛋白质是炎性细胞因子(信使)，在向血细胞发出损伤或感染位置的信号方面至关重要，因此对一生中维持健康的免疫系统至关重要。我们现在知道，对损伤敏感的“P2X7”细胞表面受体的激活会导致炎症介质(特别是“白介素1”)释放(分泌)到血液中。有趣的是，当白介素1离开细胞时，它被包裹在小的、受保护的膜包中，我们称之为“微泡”。这些包可能会阻止这种高度炎症的细胞因子在体内的随机位置发挥作用，并可能提供一种将其靶向于微泡可以对接的特定细胞的方法。协调微泡形成和细胞因子分泌的细胞机制是复杂的。基于对培养中培养的单一类型细胞的研究，已经提出了几种参与这一过程的途径。我们已经开发了一种新的系统，我们可以在整个活体中研究荧光细胞因子，即透明斑马鱼的幼体。这首次为我们提供了观察分泌白介素1的细胞的机会，并了解了白介素1在活体中的靶点。在这个项目中，我们将使用我们的荧光斑马鱼品系来优化计算方法，以量化整个生物体中囊泡的形成、跟踪和靶向。然后，我们将测试哪些细胞成分负责白介素1的分泌，方法是向这些成分添加抑制剂，并测量对感染的反应的变化。我们还将使用这些荧光斑马鱼来筛选有效阻止炎症的小分子，这样我们就可以确定细胞中健康免疫反应所需的关键部分。在未来，除了这个项目，这将帮助我们开发抗击疾病的新疗法。

项目成果

CFTR Protects against Mycobacterium abscessus Infection by Fine-Tuning Host Oxidative Defenses.

• DOI: 10.1016/j.celrep.2019.01.071
• 发表时间: 2019-02-12
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Bernut, Audrey;Dupont, Christian;Kremer, Laurent
• 通讯作者: Kremer, Laurent

Zebrafish tissue injury causes upregulation of interleukin-1 and caspase-dependent amplification of the inflammatory response.

• DOI: 10.1242/dmm.013029
• 发表时间: 2014-02
• 期刊: Disease models & mechanisms
• 影响因子: 4.3
• 作者: Ogryzko NV;Hoggett EE;Solaymani-Kohal S;Tazzyman S;Chico TJ;Renshaw SA;Wilson HL
• 通讯作者: Wilson HL

Identification of benzopyrone as a common structural feature in compounds with anti-inflammatory activity in a zebrafish phenotypic screen.

• DOI: 10.1242/dmm.024935
• 发表时间: 2016-06-01
• 期刊: Disease models & mechanisms
• 影响因子: 4.3
• 作者: Robertson AL;Ogryzko NV;Henry KM;Loynes CA;Foulkes MJ;Meloni MM;Wang X;Ford C;Jackson M;Ingham PW;Wilson HL;Farrow SN;Solari R;Flower RJ;Jones S;Whyte MK;Renshaw SA
• 通讯作者: Renshaw SA

A152T tau allele causes neurodegeneration that can be ameliorated in a zebrafish model by autophagy induction.

• DOI: 10.1093/brain/awx005
• 发表时间: 2017-04-01
• 期刊: Brain : a journal of neurology
• 作者: Lopez A;Lee SE;Wojta K;Ramos EM;Klein E;Chen J;Boxer AL;Gorno-Tempini ML;Geschwind DH;Schlotawa L;Ogryzko NV;Bigio EH;Rogalski E;Weintraub S;Mesulam MM;Tauopathy Genetics Consortium;Fleming A;Coppola G;Miller BL;Rubinsztein DC
• 通讯作者: Rubinsztein DC

Cryptococcus neoformans Intracellular Proliferation and Capsule Size Determines Early Macrophage Control of Infection.

• DOI: 10.1038/srep21489
• 发表时间: 2016-02-18
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Bojarczuk A;Miller KA;Hotham R;Lewis A;Ogryzko NV;Kamuyango AA;Frost H;Gibson RH;Stillman E;May RC;Renshaw SA;Johnston SA
• 通讯作者: Johnston SA