Functional role(s) of oestrogen signalling on neuronal progenitor cell development and fate in the brain

雌激素信号对大脑神经祖细胞发育和命运的功能作用

基本信息

批准号：
BB/L020637/1
负责人：
Charles Tyler
金额：
$ 59.51万
依托单位：
UNIVERSITY OF EXETER
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2014
资助国家：
英国
起止时间：
2014 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FL020637%2F1
关键词：
Functional role oestrogen signalling neuronal

项目摘要

Nerve cells (called neurons) form in the brain principally during embryo development but they can also be synthesised in the adult brain. Defects in the process of neuron synthesis (neurogenesis) have been linked with various brain diseases, psychiatric illnesses and addictions. Neurogenesis is also critically involved in the repair of an injured brain. One type of cell, called radial glial cells (RGCs) can give rise to all types of neurons in the cerebral cortex of the brain but how this is controlled is not well understood. Recently, hormones, called oestrogens, have been indicated to play important roles in RGCs and in neurogenesis.In this project we will apply genetically engineered zebrafish models to investigate the roles of oestrogen in neurogenesis and brain development, investigating also for sex related differences. We are using zebrafish for this work because of the relative ease to create transgenic animals and the fact that there is a high neurogenic activity in the brain throughout life. We have already successfully developed one genetically engineered (transgenic) zebrafish in which cells responsive to oestrogen produce a green fluorescent protein (GFP) that can be detected using imaging methods in living animals and in real time. Using this model we will establish the role of oestrogens in the RGCs in the telecephalon and identify the different cells in the brain responsive to oestrogens and how they develop. We will also apply a method called two-photon microscopy to selectively remove the oestrogen responsive RGCs in the telecephalon and investigate subsequent effects on brain development to help establish their function. We will then develop a new transgenic zebrafish model with a different genetic insert that prolongs the green fluorescent signal generated in response to oestrogen exposure. This will enable us to follow the fate of the oestrogen responsive RGCs and other neural cells of early life in the brain of juvenile and adult fish. We will also use this model to investigate for differences that occur between males and females.We will develop a further, more complex (triple), transgenic zebrafish that will enable us to identify activity in oestrogen responding neurons. This model includes two fluorescent colours, red identifying oestrogen responsive neurons and green identifying when these cells are active (produced in response to increased levels of calcium in the cell). Using this fish we will investigate the roles of oestrogen responding neurons in smell, including assessing for sex related differences. We will also apply a technique to block oestrogen action responsive neurons in the telecephalon during early life to establish how this affects their subsequent development and function in smell.Applying these transgenic models, finally we will initiate studies into the effects of exposure to so called endocrine disrupting chemicals (EDCs), for which there is widespread health concern, on oestrogen responding neurons and their subsequent fate and function. The research will be significant interest to a diverse audience including academic and industry researchers, and the medical profession, by provision of new models to study neurogenesis and the roles of an important group of environmental chemicals on human health. It will also be of strong interest to industry and government regulatory bodies, as the models develop could be applied for advancing risk assessment of chemicals with oestrogenic activity, supporting evidence-based decision-making for those chemicals. The wider public will benefit also from this research from improved understanding of neurological conditions and better evidence of chemical effects that may affect their own health. The models developed will provide more integrative systems for chemical effects analysis with great potential for reducing the numbers of vertebrate animals used in testing.

神经细胞（称为神经元）主要在胚胎发育过程中在大脑中形成，但它们也可以在成年大脑中合成。神经元合成（神经发生）过程中的缺陷与各种脑部疾病、精神疾病和成瘾有关。神经发生对于受损大脑的修复也至关重要。一种称为放射状胶质细胞（RGC）的细胞可以在大脑皮层中产生所有类型的神经元，但其控制方式尚不清楚。最近，雌激素已被证明在 RGC 和神经发生中发挥重要作用。在这个项目中，我们将应用基因工程斑马鱼模型来研究雌激素在神经发生和大脑发育中的作用，同时也研究性别相关的差异。我们使用斑马鱼进行这项工作，因为创造转基因动物相对容易，而且大脑在一生中都存在高度的神经源性活动。我们已经成功开发了一种基因工程（转基因）斑马鱼，其中对雌激素敏感的细胞产生绿色荧光蛋白（GFP），可以使用活体动物中的成像方法实时检测到该蛋白。使用该模型，我们将确定雌激素在端脑 RGC 中的作用，并确定大脑中对雌激素敏感的不同细胞及其发育方式。我们还将应用一种称为双光子显微镜的方法来选择性地去除端脑中的雌激素反应性 RGC，并研究随后对大脑发育的影响，以帮助确定其功能。然后，我们将开发一种新的转基因斑马鱼模型，该模型具有不同的基因插入，可以延长因雌激素暴露而产生的绿色荧光信号。这将使我们能够追踪幼鱼和成鱼大脑中雌激素反应性 RGC 和其他早期生命神经细胞的命运。我们还将使用这个模型来研究雄性和雌性之间发生的差异。我们将开发一种更复杂的（三重）转基因斑马鱼，这将使我们能够识别雌激素反应神经元的活动。该模型包括两种荧光颜色，红色用于识别雌激素反应神经元，绿色用于识别这些细胞何时活跃（响应细胞中钙水平的增加而产生）。使用这条鱼，我们将研究雌激素反应神经元在嗅觉中的作用，包括评估性别相关的差异。我们还将应用一种技术来阻断生命早期端脑中雌激素作用反应性神经元，以确定这如何影响它们随后的发育和嗅觉功能。应用这些转基因模型，最后我们将开始研究暴露于所谓的内分泌的影响干扰化学物质（EDC）对雌激素反应神经元及其随后的命运和功能产生了广泛的健康关注。该研究通过提供新模型来研究神经发生以及一组重要的环境化学物质对人类健康的作用，将引起包括学术和行业研究人员以及医学界在内的不同受众的极大兴趣。行业和政府监管机构也对此产生了浓厚的兴趣，因为开发的模型可用于推进具有雌激素活性的化学品的风险评估，支持对这些化学品的循证决策。更广泛的公众也将从这项研究中受益，因为他们可以更好地了解神经系统疾病，并获得可能影响自身健康的化学效应的更好证据。开发的模型将为化学效应分析提供更加综合的系统，在减少测试中使用的脊椎动物数量方面具有巨大的潜力。