Association of Thy-1 Differentiation Alloantigen with Synaptic Complexes Isolated from Mouse Brain

Conventional fractionation procedures were used in an effort to define the subcellular distribution of the Thy-1 alloantigen in whole mouse brain. After discontinuous sucrose density gradient centrifugation of isotonic postnuclear particulate fractions, the bulk of Thy-1 was recovered in regions of...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1978-07, Vol.75 (7), p.3283-3287
Main Authors: Acton, Ronald T., Addis, Jane, Carl, G. F., McClain, L. D., Bridgers, William F.
Format: Article
Language:English
Subjects:
Animals
Antiserum
Brain - immunology
Brain - ultrastructure
Cell lines
Cell membranes
Centrifugation
Isoantigens
Mice
Mice, Inbred Strains
Myelin
Polyethylene Glycols
Subcellular Fractions - immunology
Synaptic Membranes - immunology
Synaptosomes
Synaptosomes - immunology
T lymphocytes
Thymocytes
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Summary:Conventional fractionation procedures were used in an effort to define the subcellular distribution of the Thy-1 alloantigen in whole mouse brain. After discontinuous sucrose density gradient centrifugation of isotonic postnuclear particulate fractions, the bulk of Thy-1 was recovered in regions of the gradients containing synaptosomes. The synaptosome fraction that banded at 1.2 M sucrose yielded a specific activity for Thy-1 significantly greater than the synaptosome separating at 1.4 M sucrose. Osmotic lysis of both synaptosome fractions resulted in further enrichment in Thy-1 activity with no concomitant decrease in yields. The synaptosomal membranes obtained in this way were subsequently treated with Triton X-100 and subjected to further density gradient centrifugation. Although the detergent treatment resulted in some loss of antigenic activity, the gradient fractions that contained Thy-1 also were found by electron microscopy to be richest in synaptic junctional complexes. These findings suggest that Thy-1 is associated with synaptosomes and synaptic junctional complexes and therefore may be involved in the formation and/or maintenance of synaptic connections.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.75.7.3283