STAR Protoc. 2026 Mar 20;7(2):104448. doi: 10.1016/j.xpro.2026.104448
Protocol for preparation of a synaptosome-enriched fraction to profile presynaptic RNA content in adult Drosophila brains
Lauren Blot1, Agnès Loubat1, Marine Dehecq1, Florence Besse2
Affiliations
1Université Côte d’Azur, CNRS, Inserm, Institut de Biologie Valrose, Nice, France.
2Université Côte d’Azur, CNRS, Inserm, Institut de Biologie Valrose, Nice, France
Abstract
Neuronal synapses contain rich repertoires of RNAs that can be used for onsite translation and modulation of their proteomes. Here, we present a protocol to recover a synaptosome-enriched fraction adapted to presynaptic RNA profiling, starting from adult Drosophila heads. We describe steps for generating synaptosomes through gentle lysis, followed by centrifugation and a density-based sucrose gradient. We then detail the procedures for validation of the protocol. This protocol can be applied to frozen material. For complete details on the use and execution of this protocol, please refer to de Queiroz et al.1.