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The Specificity and Cross-Reactivity of Antisynaptosome Antibodies as Determined by Sequential Adsorption Analysis^1,2,

Department of Microbiology and Immunology and the Division of Neurology, Duke University Medical Center, Durham, North Carolina 27706

Abstract

An improved iso-osmotic, Ficoll-sucrose, density-gradient technique for isolating synaptosomes, synaptic membranes, brain mitochondria and three forms of myelin from adult rat brain is described. Antibodies raised in rabbits to isolated synaptosomes, synaptic membranes, brain mitochondria and liver mitochondria, when assayed by means of sequential adsorption analysis, showed that a major class of determinants is shared by brain mitochondria and synaptic membranes, that another major class is shared by myelin and synaptic membranes and that a third class is shared by liver mitochondria and the various brain fractions. Besides these crossreactivities synaptosomes also displayed a class of distinctive determinants separate from those of myelin; brain mitochondria displayed a class of distinctive determinants separate from those of synaptic membranes and from liver mitochondria, and liver mitochondria displayed a class of distinctive determinants separate from those of brain mitochondria. Synaptosomes also combined with a small but definite amount of antibody in two different antisera that did not cross-react either with isolated synaptic membranes or brain mitochondria at the given concentrations, thereby revealing a distinctive class of synaptosome determinants, possibly conformational in nature.

Footnotes

¹ Supported by Atomic Energy Commission Contract AT-(40-1)-3195, National Institutes of Health Grant NB-07872 from the National Institutes of Neurological Diseases and Stroke, and the Robert McManus Memorial Grant No. 558-B-3 from the National Multiple Sclerosis Society.

² Part of this work was reported at the 1971 FASEB meeting, Chicago, Ill.

³ Supported by a National Institutes of Health Post-Doctoral Traineeship, MH-08394.

⁴ Supported by a National Institutes of Health Pre-Doctoral Traineeship, 5T01-AI 00285. Part of this work appears in a dissertation submitted in partial fulfillment of the requirements for the Ph.D. degree.

⁵ Career Development Awardee of the United States Public Health Service, Division of Neurology.

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