Cytoplasmic expression of the leu-4 (CD3) antigen in developing Purkinje cells in the rat cerebellum

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Abstract
Although commonly known to represent a T cell receptor (CD3) associated polypeptide, the leu-4 (CD3) antigen occurs in cerebellar Purkinje cells (PCs) of many species. The monoclonal pan T lymphocyte marker anti-leu-4 (CD3) recognizes both the lymphocytic and the Purkinje cell type of this antigen [22]. To obtain more information about the merit of anti-leu-4 (CD3) as an investigational tool, we evaluated the expression of leu-4 (CD3) in PCs of the developing rat cerebellum (in situ) by light microscopy. Positive anti-leu-4 (CD3) immunoreaction of PCs did not occur prior to post-natal day (D) 4. The analysis of immunostaining during cell differentiation revealed three major phases of post-natal PC maturation including antigenic development of cell somata (phase 1: until D6), dendrites (phase 2: D7-D11), and axons (phase 3: D12-D14). A massive post-weaning expansion of the dendritic arborization led then to the mature PC architecture. Additionally, the leu-4 (CD3) antigen was observed in ectopic PC dendrites (D10) and in ectopic (mature) PCs. Throughout post-natal development as well as in mature PCs, the leu-4 (CD3) antigen was found to be cytoplasmic. Due to its labile nature, neither an ultrastructural localization nor molecular characterization could be achieved. For the same reason, its application is basically restricted to cryo-fixed cerebellar tissue. However, at the level of light microscopy, the monoclonal human T cell marker anti-leu-4 (CD3) proved to be a useful tool for specific and sensitive labelling of differentiated cerebellar PCs in the rat.

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