A NEW INCUBATION TECHNIQUE FOR THE STUDY OF SYNAPTOSOMAL TRANSPORT |
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| Authors: | Shing-Mei Hwang Karl S Roth Stanton Segal |
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| Institution: | Division of Biochemical Development and Molecular Diseases, Children's Hospital of Philadelphia, and Departments of Pediatrics and Medicine, University of Pennsylvania School of Medicine. Philadelphia, PA 19104, U.S.A. |
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| Abstract: | A simple, inexpensive method is described for the incubation and study of transport in synaptosomes. This method avoids the many potential sources of error inherent in the methods currently in use. The transport of two sugars and two amino acids has been studied, using this system, and the resulting data is compared to similar studies already published by others. A steady-state was achieved and maintained for 60 min independent of tissue concentration, except in the case of γ-aminobutyric acid. An analysis of the entry kinetics of 2-deoxy-d -glucose has been performed, the results indicating that the present method offers the advantage of substantially increasing oxygenation and hence, efficiency of the uptake. Using α-aminoisobutyric acid, analysis of substrate flux within the system was performed, illustrating the utility of the method. The specific advantages offered by the technique over other methods in use are discussed. It is concluded that adequate oxygenation of the synaptosome during study is essential to reliable results and can easily be achieved by means of the methodology described in this report. |
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