The Molecular Mechanisms Of Plastic Changes In The Auditory Pathway During Different Auditory Activity

Purpose To prepare highly purified synaptosomes from primary auditory cortex. Methods We used two discontinuos gradient centrifugtions to prepare highly purified synaptosomes. Results Electron microscopic analysis revealed that the fractions after two gradients mainly contained synaptosomes. In addition, contained identifiable myelin, mitochondria, and considerable unidentified membranous and granular material, synaptosomes consisted of sealed presynaptic vesicles and postsynaptic density. The fractions from discontinuos percoll sedimentation gradient was more enriched for myelin and with more intrasynaptosomal mitochondria than the fractions from discontinuous Optiprep floatation gradient and essentially free of contamination from nuclear-soma-associated factors. Western blot confirmed the presence of presynaptic structures and postsynaptic proteins can be found in both total brain extract and in synaptosomes. Conclusion This method was suitable for the purification of synaptosomes in primary auditory cortex.