一种用于棘鱼组织的荧光杂交（FISH）方案。
A fluorescence hybridization (FISH) protocol for stickleback tissue.
作者信息
James Noelle, Liu Xiaochen, Bell Alison
机构信息
Neuroscience Program, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
出版信息
Evol Ecol Res. 2016;17:603-617.
BACKGROUND
Threespine stickleback are an important model for behaviour and evolutionary studies. A growing number of quantitative trait loci (QTL) and gene expression studies are identifying genes related to ecologically important traits in sticklebacks. In order to visualize the expression of candidate genes, we developed a fluorescence hybridization (FISH) protocol.
METHODS
We present a protocol for FISH on fresh or flash-frozen dissected tissue, using either cryo- or paraffin embedding. The protocol covers probe design guidelines and synthesis, sample embedding, sectioning, and the hybridization process. The protocol is optimized for brain tissue. Key steps for modifying the protocol for other tissues are noted.
RESULTS
The FISH protocol resulted in specific labelling under all combinations of dissection and embedding conditions. Paraffin embedding preserved morphology better than cryo-embedding. We provide representative results showing the expression of glial fibrillary acidic protein (), oxytocin receptor (), and tyrosine hydroxylase () in the brain.
背景
三刺鱼是行为和进化研究的重要模型。越来越多的数量性状基因座（QTL）和基因表达研究正在确定与三刺鱼生态重要性状相关的基因。为了可视化候选基因的表达，我们开发了一种荧光杂交（FISH）方案。
方法
我们展示了一种在新鲜或速冻解剖组织上进行FISH的方案，使用冷冻或石蜡包埋。该方案涵盖探针设计指南和合成、样品包埋、切片和杂交过程。该方案针对脑组织进行了优化。记录了针对其他组织修改该方案的关键步骤。
结果
FISH方案在解剖和包埋条件的所有组合下都产生了特异性标记。石蜡包埋比冷冻包埋更好地保存了形态。我们提供了显示胶质纤维酸性蛋白（）、催产素受体（）和酪氨酸羟化酶（）在大脑中表达的代表性结果。
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