Temporal and spatial profiling of gene expression during polar growth of Aspergil

描述（由申请人提供）：侵袭性曲霉病（IA）是白血病和骨髓移植患者最常见的感染性死亡原因。烟曲霉（Aspergillus fumigatus）的分生孢子是IA最常见的病原体，被吸入肺中并在肺中萌发。与所有丝状真菌一样，A.烟曲霉通过芽管和后来的分枝经历高度极性生长。这种高极性的尖端生长允许A.烟曲霉侵入血管和组织，导致IA特征性坏死。尽管极性生长对IA的重要性，但对IA极性生长的分子机制知之甚少。烟熏。这项研究将使用微阵列分析来鉴定在A.烟熏。在极性生长期间表达的基因的时间分布将使用从同步培养物中分离的RNA产生，所述同步培养物正好在芽管和分支出现之前和之后以及在菌丝和分支伸长期间。利用激光微捕获技术从芽管、菌丝和分枝的特定区域分离的RNA，将产生极性生长过程中表达的基因的空间分布。RNA将被标记并与现有PFGRC A杂交。将分析烟曲霉微阵列和表达模式。鉴定极性生长过程中差异转录和/或不对称定位的基因将为重点研究A.烟曲霉及其在IA中的作用。 公共卫生相关性：侵袭性曲霉病（IA）是一种免疫功能低下的致命性疾病，由吸入烟曲霉孢子并在肺中生长引起。与其它丝状真菌一样，A.烟曲霉发出称为菌丝的长管状细胞进入环境中寻找营养，在IA中，正是这些菌丝导致宿主组织的破坏。这项拟议中的研究将确定允许A.烟曲霉产生菌丝，从而将发现对抗IA的新方法。

DESCRIPTION (provided by applicant): Invasive aspergillosis (IA) is the most frequent infectious cause of death in leukemia and bone marrow transplant patients. The conidia of Aspergillus fumigatus, the filamentous fungus that is the most common cause of IA, are inhaled and germinate in the lung. Like all filamentous fungi A. fumigatus undergoes highly polar growth via germ tubes and later branches. This highly polar tip growth allows A. fumigatus to invade blood vessels and tissues resulting in the necrosis characteristic of IA. Despite the importance of polar growth to IA, little is known about the molecular mechanisms of polar growth in A. fumigatus. The proposed research will use microarray profiling to identify genes that are differentially transcribed and/or asymmetrically localized during polar growth in A. fumigatus. Temporal profiles of genes expressed during polar growth will be generated using RNA isolated from synchronous cultures just before and after germ tube and branch emergence and during elongation of hyphae and branches. Spatial profiles of genes expressed during polar growth will be generated using RNA isolated from specific regions of germ tubes, hyphae and branches with laser microcapture techniques. RNA will be labeled and hybridized to existing PFGRC A. fumigatus microarrays and expression patterns will be analyzed. Identification of genes that are differentially transcribed and/or asymmetrically localized during polar growth will lay the foundation for a comprehensive research program focusing on polar growth in A. fumigatus and its role in IA. PUBLIC HEALTH RELEVANCE: Invasive aspergillosis (IA), an often fatal disease of the immunocompromised, is caused when the spores of Aspergillus fumigatus are inhaled and grow in the lung. Like other filamentous fungi, A. fumigatus sends out long tubular cells known as hyphae into the environment searching for nutrients and in IA it is these hyphae which cause the destruction of host tissue. The proposed research will identify genes that allow A. fumigatus to make hyphae and so will uncover new ways of combating IA.