Kucera Aurelia C, Heidinger Britt J
Environmental and Conservation Sciences Program, North Dakota State University;
Department of Biological Sciences, North Dakota State University.
J Vis Exp. 2018 Feb 5(132):55324. doi: 10.3791/55324.
Collection of semen may be useful for a wide range of applications including studies involving sperm quality, sperm telomere dynamics, and epigenetics. Birds are widely used subjects in biological research and are ideal for studies involving repeated sperm samples. However, few resources are currently available for those wishing to learn how to collect and extract DNA from avian sperm. Here we describe cloacal massage, a gentle, non-invasive manual technique for collecting avian sperm. Although this technique is established in the literature, it can be difficult to learn from the available descriptions. We also provide information for extracting DNA from avian semen using a commercial extraction kit with modifications. Cloacal massage can be easily used on any small- to medium-sized male bird in reproductive condition. Following collection, the semen can be used immediately for motility assays, or frozen for DNA extraction following the protocol described herein. This extraction protocol was refined for avian sperm and has been successfully used on samples collected from several passerine species (Passer domesticus, Spizella passerina, Haemorhous mexicanus, and Turdus migratorius) and one columbid (Columba livia).
采集精液可用于广泛的应用，包括涉及精子质量、精子端粒动态和表观遗传学的研究。鸟类是生物学研究中广泛使用的对象，非常适合进行涉及重复采集精子样本的研究。然而，目前对于那些希望学习如何从鸟类精子中采集和提取DNA的人来说，可用的资源很少。在此，我们描述泄殖腔按摩法，这是一种轻柔、非侵入性的手动采集鸟类精子的技术。尽管该技术在文献中有记载，但从现有的描述中可能难以学会。我们还提供了使用经过改良的商业提取试剂盒从鸟类精液中提取DNA的信息。泄殖腔按摩法可轻松用于处于繁殖状态的任何中小型雄性鸟类。采集后，精液可立即用于活力测定，或按照本文所述的方案冷冻后用于DNA提取。这种提取方案是针对鸟类精子进行优化的，已成功应用于从几种雀形目鸟类（家雀、歌带鹀、紫翅椋鸟和美洲知更鸟）和一种鸽形目鸟类（家鸽）采集的样本。
