Bostan Sevginur, Serdengeçti Safiye, Bayat F Kemal, Bay Sadık, Sezer AyşeServer, Ayşit Neşe, Öztürk Gürkan
Research Institute for Health Sciences and Technologies (SABITA), Regenerative and Restorative Medicine Research Center (REMER), Istanbul Medipol University, Istanbul, Türkiye.
Department of Physiology, Graduate School of Health Sciences, Istanbul Medipol University, Istanbul, Türkiye.
J Comp Neurol. 2025 Jun;533(6):e70066. doi: 10.1002/cne.70066.
The axolotl (Ambystoma mexicanum), a neotenic salamander with remarkable regenerative capabilities, serves as a key model for studying nervous system regeneration. Despite its potential, the cellular and molecular mechanisms underlying this regenerative capacity remain poorly understood, partly due to the lack of reliable in vitro models for axolotl neural cells. In this study, we developed a novel protocol for primary cultures of adult axolotl telencephalon/pallium, enabling the maintenance of viable and functionally active neural cells. Using calcium imaging and immunocytochemistry, we demonstrated the presence of neuronal and glial markers, synaptic connections, and spontaneous calcium activity, highlighting the functional integrity of the cultured cells. Our findings reveal that these cultures can be maintained in both serum and serum-free conditions, with neurons exhibiting robust neurite outgrowth and responsiveness to injury. This protocol addresses a critical gap in axolotl research by providing a controlled in vitro system to study neurogenesis and regeneration. By offering insights into the regenerative mechanisms of axolotl neurons, this work lays the foundation for comparative studies with mammalian systems, potentially informing therapeutic strategies for neurodegenerative diseases and CNS injuries in humans.
美西钝口螈（Ambystoma mexicanum）是一种具有显著再生能力的幼态延续蝾螈，是研究神经系统再生的关键模型。尽管它具有潜力，但这种再生能力背后的细胞和分子机制仍知之甚少，部分原因是缺乏可靠的美西钝口螈神经细胞体外模型。在本研究中，我们开发了一种用于成年美西钝口螈端脑/大脑皮层原代培养的新方案，能够维持有活力且功能活跃的神经细胞。通过钙成像和免疫细胞化学，我们证明了神经元和神经胶质细胞标志物、突触连接以及自发钙活性的存在，突出了培养细胞的功能完整性。我们的研究结果表明，这些培养物可以在有血清和无血清条件下维持，神经元表现出强大的神经突生长和对损伤的反应能力。该方案通过提供一个可控的体外系统来研究神经发生和再生，填补了美西钝口螈研究中的一个关键空白。通过深入了解美西钝口螈神经元的再生机制，这项工作为与哺乳动物系统的比较研究奠定了基础，可能为人类神经退行性疾病和中枢神经系统损伤的治疗策略提供信息。
