Isolation of nerve endings from the central nervous system of the cricket (Acheta domesticus) using a percoll gradient technique

Abstract

A method is described for the isolation of nerve endings from the central nervous system of the cricket (Acheta domesticus). The fraction is relatively free of contaminating particles and releases transmitter efficiently in response to depolarising stimuli. The procedure which employs Percoll in isoosmotic mannitol results in an average yield of 5.66 mg synaptosomes per gram wet weight and is highly reproducible. The separation technique is also reasonably rapid, taking <2h and has the advantage that a large proportion of the nerve membrane vesicles generated during homogenization and tissue fractionation effectively separate from the synaptosomal layer during centrifugation. Neuronal mitochondria can also be isolated in good purity using this procedure.