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Recently, Percoll gradients have been successfully used for the isolation of synaptosomes. Te present report describes two procedures for isolating synaptosomes using discontinous Percoll gradients. One of them uses a superspeed centrifuge (Sorvall RC 2B, fixed-angle rotor SS-34), while the other uses an ultracentrifuge swinging bucket rotor (SW 50.1). Using this swinging rotor synaptosomes can be obtained from small tissue samples (100 mg). The partial caracterization of the fractions resulting from gradients demonstrated that intact synaptosomes were recovered from the 10%/15% and 10/16% Percoll interphases for the superspeed and ultracentrifuge, respectively. The mitochondrial contamination of the synaptosomal fractions was similar in both procedures and also similar to those reported in literature. The membrane contamination of the synaptosomal fraction obtained using the swinging roto r was very low, while the use of SS-34 roto r yield a fraction more contaminated with membranes. The results of the present study demonstrate that synaptosomes can be obtained by using relatively simple centrifuges (superspeed) or from small tissue sample (swinging rotor SW 50.1). The time of preparation range from 20 to 60 minutes.

Dois procedimentos para isolar sinaptossomas usando gradientes de Percoll