Genetics and Genome Biology, School of Biological Sciences, University of Leicester, Leicester, England, LE1 7RH, UK.
Neuroscience, Psychology and Behaviour, University of Leicester, Leicester, Leicestershire, LE1 7RH, UK.
F1000Res. 2024 Jun 27;10:1064. doi: 10.12688/f1000research.73115.1. eCollection 2021.
Fish species are commonly used as experimental models in the laboratory. DNA is routinely collected from these animals to permit identification of their genotype. The current standard procedure to sample DNA is fin clipping, which involves anaesthetising individuals and removing a portion of the caudal fin. While fin clipping reliably generates good quality DNA samples for downstream applications, there is evidence that it can alter health and welfare, and impact the fish's behaviour. This in turn can result in greater variation in the data collected. In a recent study we adapted a skin swabbing protocol to collect DNA from small-bodied fish, including sticklebacks and zebrafish, without the use of analgesics, anaesthetics or sharp instruments. A rayon-tipped swab was used to collect mucus from the flank of the fish, which was then used for DNA extraction. We subsequently demonstrated that compared to fin clipping, skin swabbing triggered fewer changes in stress axis activation and behaviour. We also found that gene expression and behaviour data collected from swabbed fish were less variable than similar data collected from fish that had been fin clipped. This potentially allows smaller sample sizes in experimental groups to be used after skin swabbing, thereby reducing animal use. Here we provide a detailed protocol explaining how to collect DNA samples from small laboratory fish using skin swabs.
鱼类通常被用作实验室的实验模型。为了允许鉴定它们的基因型,通常会从这些动物身上采集 DNA。目前,从鱼类身上采集 DNA 的标准程序是剪鳍,涉及麻醉个体并切除一部分尾鳍。虽然剪鳍可以可靠地为下游应用生成高质量的 DNA 样本,但有证据表明它会改变鱼类的健康和福利,并影响其行为。这反过来又会导致收集到的数据有更大的差异。在最近的一项研究中,我们改编了一种皮肤擦拭协议,无需使用镇痛剂、麻醉剂或尖锐仪器,即可从小体型鱼类(包括刺鱼和斑马鱼)身上采集 DNA。使用粘胶纤维拭子从鱼的侧腹收集黏液,然后用于 DNA 提取。我们随后证明,与剪鳍相比,皮肤擦拭对应激轴激活和行为的影响较小。我们还发现,与从剪鳍鱼身上采集的类似数据相比,从擦拭过的鱼身上采集的基因表达和行为数据的变异性更小。这可能允许在皮肤擦拭后,在实验组中使用更小的样本量,从而减少动物的使用。在这里,我们提供了一个详细的协议,说明如何使用皮肤拭子从小型实验室鱼类中采集 DNA 样本。