Marine mammals exhibit some of the most dramatic physiological adaptations in their clade and offer unparalleled insights into the mechanisms driving convergent evolution on relatively short time scales. Some of these adaptations, such as extreme tolerance to hypoxia and prolonged food deprivation, are uncommon among most terrestrial mammals and challenge established metabolic principles of supply and demand balance. Non-targeted omics studies are starting to uncover the genetic foundations of such adaptations, but tools for testing functional significance in these animals are currently lacking. Cellular modeling with primary cells represents a powerful approach for elucidating the molecular etiology of physiological adaptation, a critical step in accelerating genome-to-phenome studies in organisms in which transgenesis is impossible (e.g., large-bodied, long-lived, fully aquatic, federally protected species). Gene perturbation studies in primary cells can directly evaluate whether specific mutations, gene loss or duplication confer functional advantages such as hypoxia or stress tolerance in marine mammals. Here, we summarize how genetic and pharmacological manipulation approaches in primary cells have advanced mechanistic investigations in other non-traditional mammalian species, and highlight the need for such investigations in marine mammals. We also provide key considerations for isolating, culturing, and conducting experiments with marine mammal cells under conditions that mimic in vivo states. We propose that primary cell culture is a critical tool for conducting functional mechanistic studies (e.g. gene knockdown, over-expression or editing) that can provide the missing link between genome- and organismal-level understanding of physiological adaptations in marine mammals.

中文翻译：

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原代细胞的功能研究为海洋哺乳动物的基因组到表型组研究提供了一个系统

海洋哺乳动物在其进化枝中表现出一些最显着的生理适应，并为在相对较短的时间尺度上驱动趋同进化的机制提供了无与伦比的见解。其中一些适应，例如对缺氧的极端耐受和长时间的食物剥夺，在大多数陆生哺乳动物中并不常见，并且挑战了既定的供需平衡代谢原则。非靶向组学研究开始揭示这种适应的遗传基础，但目前缺乏在这些动物中测试功能意义的工具。用原代细胞进行细胞建模代表了阐明生理适应的分子病因学的有力方法，这是加速生物体基因组到现象研究的关键步骤，其中转基因是不可能的（例如，体大、寿命长、完全水生、受联邦保护的物种）。原代细胞中的基因扰动研究可以直接评估特定突变、基因丢失或重复是否赋予海洋哺乳动物缺氧或应激耐受等功能优势。在这里，我们总结了原代细胞中的遗传和药理学操作方法如何在其他非传统哺乳动物物种中进行了先进的机械研究，并强调了对海洋哺乳动物进行此类研究的必要性。我们还提供了在模拟体内状态的条件下分离、培养和进行海洋哺乳动物细胞实验的关键考虑因素。我们建议原代细胞培养是进行功能机制研究（例如基因敲除、