Background

Investigating brain function requires tools and techniques to visualise, modify and manipulate neuronal tissue. One powerful and popular method is intracerebral injection of customised viruses, allowing expression of exogenous transgenes. This technique is a standard procedure for adult mice, and is used by laboratories worldwide. Use of neonatal animals in scientific research allows investigation of developing tissues and enables long-term study of cell populations. However, procedures on neonatal mice are more challenging, due to the lack of reliable methods and apparatus for anaesthesia of these animals.

New method

Here, we report an inhalation-based protocol for anaesthesia of neonatal (P0-2) mice and present a custom 3D-printed apparatus for maintenance of anaesthesia during surgical procedures. Our optimised method of anaesthesia enables a rapid method of stereotactic injection in neonatal mice for transduction of brain tissue.

Results and comparison with existing methods

This approach significantly enhances animal welfare and facilitates wider and simpler use of neonatal rodents in scientific research. We demonstrate this procedure for targeted labelling of specific brain regions, and in vivo modification of tissue prior to organotypic culture.

Conclusions

Our protocol for reliable delivery of inhalational anaesthetics can be readily adopted by any laboratory and will enable safer use of neonatal rodents across a diverse spectrum of scientific disciplines. Application to stereotactic injections allows a rapid and efficient method for modification of brain tissue.

中文翻译：

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一种用于新生小鼠的吸入麻醉方法，允许在大脑中进行流线型立体定向注射。

背景

研究大脑功能需要工具和技术来可视化、修改和操纵神经元组织。一种强大而流行的方法是脑内注射定制病毒，允许表达外源转基因。这种技术是成年小鼠的标准程序，被世界各地的实验室使用。在科学研究中使用新生动物可以研究发育中的组织，并可以对细胞群进行长期研究。然而，由于缺乏可靠的麻醉方法和设备，对新生小鼠的操作更具挑战性。

新方法

在这里，我们报告了一种基于吸入的新生 (P0-2) 小鼠麻醉协议，并展示了一种定制的 3D 打印设备，用于在手术过程中维持麻醉。我们优化的麻醉方法使新生小鼠能够快速进行立体定向注射以转导脑组织。

结果和与现有方法的比较

这种方法显着提高了动物福利，并有助于在科学研究中更广泛、更简单地使用新生啮齿动物。我们演示了此过程，用于特定大脑区域的靶向标记，以及器官型培养之前组织的体内修饰。

结论

我们可靠地输送吸入麻醉剂的协议可以很容易地被任何实验室采用，并将使在各种科学学科中更安全地使用新生啮齿动物。应用于立体定向注射允许快速有效的方法来修改脑组织。