Thangamani Saravanan, Wikel Stephen K
Department of Pathology and Center for Biodefense and Emerging Infectious, Diseases, University of Texas Medical Branch, Galveston, Texas 77555, USA.
Parasit Vectors. 2009 Jul 24;2(1):34. doi: 10.1186/1756-3305-2-34.
Saliva of Aedes aegypti contains a complex array of proteins essential for both blood feeding and pathogen transmission. A large numbers of those proteins are classified as unknown in regard to their function(s). Understanding the dynamic interactions at the mosquito-host interface can be achieved in part by characterizing mosquito salivary gland gene expression relative to blood feeding. Towards this end, we developed an oligonucleotide microarray representing 463 transcripts to determine differential regulation of salivary gland genes. This microarray was used to investigate the temporal gene expression pattern of Ae. aegypti salivary gland transcriptome at different times post-blood feeding. Expression of the majority of salivary gland genes (77-87%) did not change significantly as a result of blood feeding, while 8 to 20% of genes were down-regulated and 2.8 to 11.6% genes were up-regulated. Up-regulated genes included defensins, mucins and other immune related proteins. Odorant-binding protein was significantly down-regulated. Among unknown function proteins, several were up-regulated during the first three hours post-blood feeding and one was significantly down-regulated. Quantitative real-time RT-PCR was used to substantiate differential expression patterns of five randomly selected genes. Linear regression analysis revealed a high degree of correlation (R2 > 0.89) between oligonucleotide microarray and quantitative RT-PCR data. To our knowledge, this is the first study to investigate differential expression of the Ae. aegypti salivary gland transcriptome upon blood feeding. A microarray provides a robust, sensitive way to investigate differential regulation of mosquito salivary gland genes.
埃及伊蚊的唾液含有一系列复杂的蛋白质，这些蛋白质对于吸血和病原体传播都至关重要。其中大量蛋白质的功能尚属未知。通过表征蚊子唾液腺相对于吸血的基因表达，能够部分地了解蚊子与宿主界面的动态相互作用。为此，我们开发了一种代表463个转录本的寡核苷酸微阵列，以确定唾液腺基因的差异调控。该微阵列用于研究埃及伊蚊唾液腺转录组在吸血后不同时间的时间基因表达模式。大多数唾液腺基因（77 - 87%）的表达并未因吸血而发生显著变化，而8%至20%的基因被下调，2.8%至11.6%的基因被上调。上调的基因包括防御素、粘蛋白和其他免疫相关蛋白。气味结合蛋白被显著下调。在功能未知的蛋白质中，有几种在吸血后的前三小时内被上调，一种被显著下调。使用定量实时RT - PCR来证实随机选择的五个基因的差异表达模式。线性回归分析显示寡核苷酸微阵列和定量RT - PCR数据之间具有高度相关性（R2 > 0.89）。据我们所知，这是第一项研究埃及伊蚊唾液腺转录组在吸血后差异表达的研究。微阵列为研究蚊子唾液腺基因的差异调控提供了一种强大、灵敏的方法。
