A fluorescence in situ hybridization (FISH) protocol for stickleback tissue.

Q2 Agricultural and Biological Sciences

Evolutionary Ecology Research Pub Date : 2016-01-01

Noelle James, Xiaochen Liu, Alison Bell

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Abstract

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 in situ 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 (GFAP), oxytocin receptor (OXTR), and tyrosine hydroxylase (TH) in the brain.

Abstract Image

本刊更多论文

棘鱼组织的荧光原位杂交(FISH)方法。

背景:三刺棘鱼是行为和进化研究的重要模型。越来越多的数量性状位点(QTL)和基因表达研究正在确定与棘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原位杂交(FISH)方案。方法:我们提出了一种在新鲜或快速冷冻的解剖组织上进行FISH的方案，使用冷冻或石蜡包埋。该协议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。指出了修改其他组织的方案的关键步骤。结果:FISH方案在所有剥离和包埋条件组合下均产生特异性标记。石蜡包埋比冷冻包埋更能保存形态学。我们提供了脑内胶质纤维酸性蛋白(GFAP)、催产素受体(OXTR)和酪氨酸羟化酶(TH)表达的代表性结果。

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来源期刊

Evolutionary Ecology Research 生物-进化生物学

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Evolutionary Ecology Research publishes original research contributions focusing on the overlap between ecology and evolution. Papers may treat any taxon or be general. They may be empirical, theoretical or a combination of the two. EER prefers conceptual contributions that take intellectual risks or that test ideas.