Clemons Anthony, Haugen Morgan, Flannery Ellen, Kast Kristopher, Jacowski Caitlin, Severson David, Duman-Scheel Molly
Department of Medical and Molecular Genetics, Indiana University School of Medicine, South Bend, Indiana 46617, USA.
Cold Spring Harb Protoc. 2010 Oct 1;2010(10):pdb.prot5508. doi: 10.1101/pdb.prot5508.
Blood-feeding mosquitoes, including the dengue and yellow fever vector Aedes aegypti, transmit many of the world's deadliest diseases. Such diseases have resurged in developing countries and pose clear threats for epidemic outbreaks in developed countries. Recent mosquito genome projects have stimulated interest in the potential for arthropod-borne disease control by genetic manipulation of vector insects, and genes that regulate development are of particular interest. This protocol describes a method for fixation and dissection of Ae. aegypti embryos, larvae, and pupae. Tissue processed in this manner can be used subsequently for in situ hybridization detection of mRNA or immunohistochemical analysis of protein expression.
吸食血液的蚊子，包括登革热和黄热病的传播媒介埃及伊蚊，传播着世界上许多最致命的疾病。这些疾病在发展中国家死灰复燃，并对发达国家的疫情爆发构成明显威胁。最近的蚊子基因组计划激发了人们对通过基因操纵病媒昆虫来控制节肢动物传播疾病的潜力的兴趣，而调控发育的基因尤其受到关注。本方案描述了一种固定和解剖埃及伊蚊胚胎、幼虫和蛹的方法。以这种方式处理的组织随后可用于mRNA的原位杂交检测或蛋白质表达的免疫组织化学分析。
