Woods Hole Center for Oceans and Human Health

项目摘要 本附录中提出的研究验证了发育软骨藻酸(DA)的假设。 暴露会影响小胶质细胞，小胶质细胞是大脑中常驻的免疫细胞，导致功能改变， 对神经元健康和神经行为缺陷的影响。小胶质细胞在脑出血中发挥重要作用 大脑中的炎症和对免疫挑战的反应。发育中的小胶质细胞功能障碍 可能由暴露在环境化学物质中引起，并可能导致大脑结构的变化， 功能和神经行为。早期生命对小胶质细胞的影响可能会产生深远的影响，包括 对后续应激源的敏感度提高，导致神经炎症增加，最终 神经退行性变。尽管这种细胞类型很重要，但人们对其作用知之甚少。 小胶质细胞在调节包括DA在内的有害藻毒素的影响中的作用。拟议的研究将 研究发育期DA对小胶质细胞激活的影响，并评估早期DA对小胶质细胞激活的影响 生命炎症对神经元存活和学习的影响。在具体目标1中，我们检验假设 发育中的DA暴露导致斑马鱼脑内小胶质细胞的激活。我们将利用共焦 成像和转基因斑马鱼表达细胞特异性荧光标记以评估细胞的变化 小胶质细胞形态显示激活。这项研究还将确定易感窗口。在……里面 具体目标2，我们将调查发育暴露对晚年生活敏感性的影响 侮辱。我们将检验这一假设，即发育过程中暴露于DA会导致 炎症和神经退行性变，以及神经行为变化，对暴露于 成鱼。这一目标将利用联想学习的良好特征的行为分析和 神经炎症和神经变性的组织学标志物。在具体目标3中，我们将测试 发育性DA暴露导致基因表达改变和上调的假说 小胶质细胞中的炎性因子。我们将描述小胶质细胞特异的转录图谱。 使用荧光激活细胞分选和RNA测序与DA暴露相关。 这项拟议研究的结果将提供有关小胶质细胞作为一种 软骨藻酸的潜在目标，以及发育暴露的风险。

Project Summary The research proposed in this supplement tests the hypothesis that developmental domoic acid (DA) exposure affects microglia, the resident immune cells in the brain, resulting in altered function, impacts on neuron health, and neurobehavioral deficits. Microglia play an important role in inflammation and responding to immune challenge in the brain. Microglial dysfunction in development can be caused by exposure to environmental chemicals and can lead to changes in brain structure, function, and neurobehavior. Early life impacts on microglia can have far-reaching impacts, including increased sensitivity to subsequent stressors causing elevated neuroinflammation and eventual neurodegeneration. Despite the importance of this cell type, very little is known about the role of microglia in mediating the effects of harmful algal toxins, including DA. The proposed research will investigate the impacts of developmental DA on microglial activation and assess the impacts of early life inflammation on neuronal survival and learning. In specific aim 1, we test the hypothesis that developmental DA exposure causes microglial activation in zebrafish brain. We will utilize confocal imaging and transgenic zebrafish expressing cell-specific fluorescent markers to assess changes in microglial morphology indicative of activation. This study will also identify windows of susceptibility. In specific aim 2, we will investigate the impacts of developmental exposure on sensitivity to later life insults. We will test the hypothesis that developmental exposure to DA will cause increased inflammation and neurodegeneration, as well as neurobehavioral changes, in response to exposure in adult fish. This aim will utilize well-characterized behavioral assays of associative learning and histological markers of neuroinflammation and neurodegeneration. In specific aim 3, we will test the hypothesis that developmental DA exposure causes altered gene expression and upregulation of inflammatory factors in microglia. We will characterize the microglia-specific transcriptional profiles associated with DA exposure using fluorescence-activated cell sorting followed by RNA sequencing. The results from the proposed study will provide important information about the role of microglia as a potential target of domoic acid, and the risks of developmental exposure.