The subcellular fractionation of the electric organ of Torpedo

Summary The acetylcholine-rich electric organ of Torpedo has been submitted to subcellular fractionation in an attempt to isolate nerve endings and synaptic vesicles derived from cholinergic neurones. Fractions containing small vesicles and granules as their only morphologically identifiable components also contained appreciable amounts of bound acetylcholine; however, it was not possible to demonstrate a specific enrichment of any one fraction with respect to bound acetylcholine as has been possible in brain. The tissue proved difficult to homogenize and few detached nerve endings (synaptosomes) were formed. A low-speed fraction rich in Na, K- activated adenosine triphosphatase contained numerous membrane fragments with tubular appendages derived from the non-innervated surface of electroplaques. Homogenization in media isotonic with elasmobranch plasma (e.g. 0.5 M sucrose + 0.33 M urea) was essential to preserve the structure of osmotically sensitive organelles (e.g. mitochondria).We wish to express our gratitude to Dr. R. D. Keynes who arranged the supply of Torpedos and to Mr G. H. C. Dowe and Miss L. Swales for skilled technical assistance. The electron microscopic facilities were provided by a grant from the Wellcome Trust and the work was supported by a grant no. NB-03928-02 (to V.P.W.) from the National Institute of Neurological Diseases and Blindness, U.S. Public Health Service. During the period of this work Dr. Sheridan held a Postdoctoral Fellowship of the U.S. Public Health Service and Dr. Israël was an Exchange Scholar of the Medical Research Council.We are also most grateful to Professor Sir Bryan Matthews, C.B.E., Sc. D., F.R.S., for providing aquarium facilities in the Physiological Laboratory of Cambridge University.