Ramos Diane, Monteiro Antonia
Department of Biological Sciences, University at Buffalo, USA.
J Vis Exp. 2007(4):208. doi: 10.3791/208. Epub 2007 May 28.
Here we present, in video format, a protocol for in situ hybridizations in pupal wings of the butterfly Bicyclus anynana using riboprobes. In situ hybridizations, a mainstay of developmental biology, are useful to study the spatial and temporal patterns of gene expression in developing tissues at the level of transcription. If antibodies that target the protein products of gene transcription have not yet been developed, and/or there are multiple gene copies of a particular protein in the genome that cannot be differentiated using available antibodies, in situs can be used instead. While an in situ technique for larval wing discs has been available to the butterfly community for several years, the current protocol has been optimized for the larger and more fragile pupal wings.
在此，我们以视频形式展示了一种使用核糖探针在蝴蝶小弄蝶蛹翅中进行原位杂交的方案。原位杂交是发育生物学的一项主要技术，有助于在转录水平研究发育组织中基因表达的时空模式。如果尚未开发出针对基因转录蛋白产物的抗体，和/或基因组中特定蛋白质有多个基因拷贝，而现有抗体无法区分这些拷贝，那么可以改用原位杂交技术。虽然蝴蝶研究群体已经有一项针对幼虫翅盘的原位技术多年，但目前的方案已针对更大且更脆弱的蛹翅进行了优化。
