The widespread use of rodents in neuroscience has prompted the development of optimized viral variants for transduction of brain cells, in vivo. However, many of the viruses developed are less efficient in other model organisms, with birds being among the most resistant to transduction by current viral tools. Resultantly, the use of genetically-encoded tools and methods in avian species is markedly lower than in rodents; likely holding the field back. We sought to bridge this gap by developing custom viruses towards the transduction of brain cells of the Japanese quail. We first develop a protocol for culturing primary neurons and glia from quail embryos, followed by characterization of cultures via immunostaining, single cell mRNA sequencing, patch clamp electrophysiology and calcium imaging. We then leveraged the cultures for the rapid screening of various viruses, only to find that all yielded poor to no infection of cells in vitro. However, few infected neurons were obtained by AAV1 and AAV2. Scrutiny of the sequence of the AAV receptor found in quails led us to rationally design a custom-made AAV variant (AAV1-T593K; AAV1*) that exhibits improved transduction efficiencies in vitro and in vivo (14- and five-fold, respectively). Together, we present unique culturing method, transcriptomic profiles of quail’s brain cells and a custom-tailored AAV1 for transduction of quail neurons in vitro and in vivo.

啮齿类动物在神经科学中的广泛使用促使开发了用于体内脑细胞转导的优化病毒变体。然而，许多开发的病毒在其他模型生物体中的效率较低，鸟类是对当前病毒工具转导最有抵抗力的动物之一。结果，在鸟类物种中使用基因编码工具和方法的情况明显少于在啮齿动物中的使用；可能会阻止该领域。我们试图通过开发针对日本鹌鹑脑细胞转导的定制病毒来弥合这一差距。我们首先开发了一种从鹌鹑胚胎中培养原代神经元和神经胶质细胞的方案，然后通过免疫染色、单细胞 mRNA 测序、膜片钳电生理学和钙成像对培养物进行表征。然后，我们利用这些培养物快速筛选各种病毒，结果发现在体外所有细胞的感染率都很低甚至没有。然而，AAV1 和 AAV2 获得的感染神经元很少。对鹌鹑中发现的 AAV 受体序列的仔细研究促使我们合理地设计了一种定制的 AAV 变体（AAV1-T593K；AAV1*），该变体在体外和体内表现出更高的转导效率（分别为 14 倍和 5 倍） . 我们一起展示了独特的培养方法、鹌鹑脑细胞的转录组学概况和用于体外和体内转导鹌鹑神经元的定制 AAV1。对鹌鹑中发现的 AAV 受体序列的仔细研究促使我们合理地设计了一种定制的 AAV 变体（AAV1-T593K；AAV1*），该变体在体外和体内表现出更高的转导效率（分别为 14 倍和 5 倍） . 我们一起展示了独特的培养方法、鹌鹑脑细胞的转录组学概况和用于体外和体内转导鹌鹑神经元的定制 AAV1。对鹌鹑中发现的 AAV 受体序列的仔细研究促使我们合理地设计了一种定制的 AAV 变体（AAV1-T593K；AAV1*），该变体在体外和体内表现出更高的转导效率（分别为 14 倍和 5 倍） . 我们一起展示了独特的培养方法、鹌鹑脑细胞的转录组学概况和用于体外和体内转导鹌鹑神经元的定制 AAV1。