Sakamoto Hirotaka, Inutsuka Ayumu
Department of Biology, Faculty of Environmental, Life, Natural Science and Technology, Okayama University, Okayama, Japan.
Ushimado Marine Institute (UMI), Faculty of Environmental, Life, Natural Science and Technology, Okayama University, Okayama, Japan.
Acta Histochem Cytochem. 2024 Apr 25;57(2):85-88. doi: 10.1267/ahc.24-00001. Epub 2024 Apr 4.
Recent advances in viral vector technology, specifically using adeno-associated virus (AAV) vectors, have significantly expanded possibilities in neuronal tracing. We have utilized the Cre/loxP system in combination with AAV techniques in rats to explore the subcellular localization of palmitoylation signal-tagged GFP (palGFP) in oxytocin-producing neurosecretory neurons. A distinctive branching pattern of single axons was observed at the level of the terminals in the posterior pituitary. Despite challenges in detecting palGFP signals by fluorescent microscopy, immunoelectron microscopy demonstrated predominant localization on the plasma membrane, with a minor presence on the neurosecretory vesicle membrane. These findings suggest that membrane-anchored palGFP may undergo exocytosis, translocating from the plasma membrane to the neurosecretory vesicle membrane. In this study, we observed characteristic axon terminal structures in the posterior pituitary of oxytocin neurons. This study indicates the importance of understanding the plasma membrane-specific sorting system in neuronal membrane migration and encourages future studies on the underlying mechanisms.
病毒载体技术，特别是使用腺相关病毒（AAV）载体的技术，最近取得的进展显著扩展了神经元追踪的可能性。我们在大鼠中利用Cre/loxP系统与AAV技术相结合，探索在产生催产素的神经分泌神经元中棕榈酰化信号标记的绿色荧光蛋白（palGFP）的亚细胞定位。在垂体后叶的终末水平观察到单个轴突独特的分支模式。尽管通过荧光显微镜检测palGFP信号存在挑战，但免疫电子显微镜显示其主要定位于质膜，在神经分泌囊泡膜上少量存在。这些发现表明膜锚定的palGFP可能经历胞吐作用，从质膜转移到神经分泌囊泡膜。在本研究中，我们观察到催产素神经元垂体后叶中特征性的轴突终末结构。本研究表明了解质膜特异性分选系统在神经元膜迁移中的重要性，并鼓励未来对潜在机制进行研究。
