Functional genomics of the enigmatic stellate cell

神秘星状细胞的功能基因组学

• 批准号: BB/L002647/1
• 负责人: Shireen Davies
• 金额: $ 54.9万
• 依托单位: University of Glasgow
• 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%2FL002647%2F1
• 关键词: Functional; genomics; enigmatic; stellate; cell

Insects comprise the most species on earth, occupying almost all environmental niches, from arctic, desert to aquatic. Harmful insects incur enormous health and economic costs - crop damage, insect-borne plant and animal diseases, cause the loss of 20% of GDP worldwide. There is increasing pressure for insect control, given insect resistance to all insecticides and increasing environmental concerns, so understanding mechanisms of insect survival and environmental tolerance may be key to novel routes for insect control. For all insects, survival depends on osmoregulation and fluid balance (homeostasis), which also allows them to withstand desiccation in low humidity environments, including that of the UK. The insect 'kidneys' - Malpighian tubules - are key tissues for fluid homeostasis. Over some years, we have investigated and established cell signalling, ion transport, functional genomics and integrative physiology in the genetically tractable D. melanogaster Malpighian tubule, which is an excellent model for insect tubules, especially Dipteran species. Examples of Dipteran insect pests relevant to the UK economy are the wheat bulb fly and the cabbage root fly (crop pests); the midge (blue-tongue, animal health); and increasingly, mosquitoes (human health). Here, we plan to understand the mechanisms and neuroendocrine control of tubule fluid secretion and responses, specifically by the tubule stellate cell; and by chloride and water transport through the stellate cell under normal and desiccation conditions. The proposed programme of work will include isolation of actively transcribed genes (the 'translatome') from specific tubule cell-types (principal and stellate cells) under normal and desiccation conditions, using a combination of a novel cell-specific translatome isolation method, as well as gene arrays (microarrays). This will allow us to assign genes to either principal or stellate cells and so understand the gene signature of each cell type; to identify cell-specific genes which are actively transcribed in response to desiccation stress; and to define those genes implicated in desiccation tolerance by also assessing the role of identified genes in whole fly desiccation tolerance assays. We also plan to define the mechanism of chloride and water flux through the stellate cells for fluid secretion and fluid homeostasis under normal and desiccation conditions; and also in response to neurohormone control. In order to do this, we will define the role of chloride channels (CLCs) and aquaporins specific to the stellate cell at the molecular and physiological level using transgenic flies to knock-down specific CLC and aquaporin genes in only stellate cells; tubule fluid secretion assays; molecular methods; bioimaging using novel transgenic reporters; cell biology; physiological measurements (this in collaboration with a US group expert in ion channel electrophysiology); and whole fly desiccation tolerance assays. Together, this will provide the first comprehensive understanding of chloride and water transport in fluid homeostasis, as well as insights into desiccation tolerance, which may in time, help identify, new, greener insecticides that target only a subset of insects.

昆虫构成了地球上最多的物种，几乎占据了从北极、沙漠到水生生物的所有环境生态位。有害昆虫造成巨大的健康和经济代价--农作物受损、虫媒动植物疾病，导致全球国内生产总值损失20%。由于昆虫对所有杀虫剂的抗药性和对环境的日益关注，昆虫控制的压力越来越大，因此了解昆虫的生存机制和环境耐受性可能是昆虫控制新途径的关键。对于所有昆虫来说，生存依赖于渗透调节和液体平衡(动态平衡)，这也使它们能够承受低湿度环境中的干燥，包括英国的环境。昆虫的“肾脏”--马氏管--是维持体液平衡的关键组织。多年来，我们已经研究并建立了遗传易感的黑腹毛虫马氏小管的细胞信号、离子转运、功能基因组学和综合生理学，这是研究昆虫小管，特别是双翅目物种的一个很好的模型。与英国经济相关的双翅目害虫有麦鳞蝇和卷心菜根蝇(农作物害虫)；蚊子(蓝舌虫，动物健康)；以及越来越多的蚊子(人类健康)。在这里，我们计划了解小管液的分泌和反应的机制和神经内分泌控制，特别是通过小管星状细胞；以及在正常和干燥条件下通过星状细胞的氯和水运输。拟议的工作方案将包括在正常和干燥条件下从特定的小管细胞类型(主细胞和星状细胞)中分离活跃转录的基因(‘翻译组’)，使用一种新的细胞特异性翻译组分离方法以及基因阵列(微阵列)。这将使我们能够将基因分配给主细胞或星状细胞，从而了解每种细胞类型的基因特征；识别对脱水胁迫反应活跃的细胞特异性基因；并通过评估所识别的基因在整个果蝇脱水耐受性分析中的作用来确定那些与脱水耐受性有关的基因。我们还计划确定在正常和干燥条件下，氯化物和水通量通过星状细胞的液体分泌和液体动态平衡的机制，以及对神经激素控制的反应。为此，我们将在分子和生理水平上确定星状细胞特有的氯离子通道(CLCs)和水通道蛋白的作用，方法包括：转基因果蝇敲除星状细胞中特定的CLC和水通道蛋白基因；管状液分泌分析；分子方法；使用新型转基因记者进行生物成像；细胞生物学；生理测量(与美国离子通道电生理学小组专家合作)；以及全蝇干燥耐受性分析。总而言之，这将首次全面了解氯化物和水在液体动态平衡中的运输，以及对干燥耐受性的洞察，这可能有助于及时发现只针对一小部分昆虫的新的、更绿色的杀虫剂。

项目成果

When two cells are better than one: specialized stellate cells provide a privileged route for uniquely rapid water flux in Drosophila renal tubule

当两个细胞比一个细胞更好时：特化的星状细胞为果蝇肾小管中独特的快速水通量提供了一条特权途径

• DOI: 10.1101/763664
• 发表时间: 2019
• 作者: Cabrero P

The corticotropin-releasing factor-like diuretic hormone 44 (DH44) and kinin neuropeptides modulate desiccation and starvation tolerance in Drosophila melanogaster.

• DOI: 10.1016/j.peptides.2016.02.004
• 发表时间: 2016-06
• 期刊: Peptides
• 影响因子: 3
• 作者: Cannell E;Dornan AJ;Halberg KA;Terhzaz S;Dow JAT;Davies SA
• 通讯作者: Davies SA

What is an epitheliome, anyway?

到底什么是上皮组？

• DOI: 10.1016/j.ibmb.2015.10.013
• 发表时间: 2015
• 期刊: Insect biochemistry and molecular biology
• 影响因子: 3.8
• 作者: Dow JA

Tracing the evolutionary origins of insect renal function.

• DOI: 10.1038/ncomms7800
• 发表时间: 2015-04-21
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Halberg, Kenneth A.;Terhzaz, Selim;Cabrero, Pablo;Davies, Shireen A.;Dow, Julian A. T.
• 通讯作者: Dow, Julian A. T.

The cell adhesion molecule Fasciclin2 regulates brush border length and organization in Drosophila renal tubules.

• DOI: 10.1038/ncomms11266
• 发表时间: 2016-04-13
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Halberg KA;Rainey SM;Veland IR;Neuert H;Dornan AJ;Klämbt C;Davies SA;Dow JA
• 通讯作者: Dow JA