Gualtieri Claudia, Vonhoff Fernando J
Department of Biological Sciences, University of Maryland Baltimore County (UMBC), Baltimore, MD, USA.
Methods Mol Biol. 2025;2910:253-262. doi: 10.1007/978-1-0716-4446-1_15.
The flow of information within the nervous system occurs via precise connections between synaptic partners. In recent years, the development of various methods to visualize synaptic contacts has helped elucidate the connectivity within complex neuronal networks. One such method is the GRASP (GFP Reconstitution Across Synaptic Partners) technique that consists of the expression of a portion of the green fluorescent protein (GFP) at each side of the synapse, allowing the reconstitution of green fluorescence depending on the proximity of the cells expressing such tools. In Drosophila, various studies have shown the successful application of GRASP in adult flies to identify synaptic partners, whereas its use at earlier stages such as in first instar larval stages remains less common. Therefore, we provide here a detailed protocol for the visualization of GRASP-based neuronal contacts within previously established synaptic partners in first and third instar larvae.
神经系统内的信息流动通过突触伙伴之间的精确连接发生。近年来，各种可视化突触接触的方法的发展有助于阐明复杂神经元网络内的连接性。一种这样的方法是GRASP（跨突触伙伴的GFP重组）技术，该技术包括在突触两侧表达绿色荧光蛋白（GFP）的一部分，根据表达此类工具的细胞的接近程度允许绿色荧光的重组。在果蝇中，各种研究表明GRASP在成年果蝇中成功应用于识别突触伙伴，而在更早阶段如第一龄幼虫阶段使用它仍然不太常见。因此，我们在此提供一个详细的方案，用于在第一龄和第三龄幼虫中先前建立的突触伙伴内可视化基于GRASP的神经元接触。
