Synaptosomes Purification

Evans, D., Williams, R.S., Shone, C.C., Hambleton, P., Melling, J. and Dolly, J.O., Botulinum neurotoxin type B. Its purification, radioiodination and interaction with rat-brain synaptosomal membranes, Eur. I. Biochem., 154, 409-416, 1986. 

Spokes, J.W., Dolly, J.O. (1980). Complete purification of beta-hungarotoxin. Characterization of its action and that of tityustoxin on synaptosomal accumulation and release of acetylcholine. Biochim. Biophys. Acta 596 81-93. 

Transmitter uptake into secretory vesicles is an ATP-dependent process. So far these studies have been restricted to isolated secretory vesicles, where intracellular substances necessary for regulation may be lost during purification. Neuroendocrine cells or synaptosomes permeabilized with both pore-forming toxins can be used to study the regulation of transmitter storage without the necessity of purifying secretory vesicles. 

Complete purification and characterization of its action on synaptosomal accumulation and release of acetylcholine 1. W, Spokes, J, O. Dolly, Biochim. Biophys. Acta 596, 81 (1980). 0-BuTX is a pre-synaptic neurotoxin that prevents acetylcholine release at skeletal neuromuscular junctions without affecting the sensitivity of the post-synaptic membrane. Its proposed phospholipase A2 activity is believed to be responsible for some of its effects at motor nerve terminals. Activity studies P. N. Strong el a l, Proc Nat,... 

Scheer, H., and Meldolesi, J. (1985). Purification of the Putative a-latrotoxin Receptor from Bovine Synaptosomal Membranes in an Active Binding Form, EMBO J. 4 323 327. This protocol is incomplete (Scheer incubates the wells for 2 h at 4° C) with radioactive a-latrotoxin. He also fails to tell how the radioactivity gets out of the wells, but with 150 [tl 0.1 M NaOH this should be no problem. 

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