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The Journal of Neuroscience, September 15, 1998, 18(18):7047-7060

Identification of a Survival-Promoting Peptide in Medium Conditioned by Oxidatively Stressed Cell Lines of Nervous System Origin

Timothy J. Cunningham¹, Lisa Hodge¹, David Speicher², Dave Reim², Carla Tyler-Polsz¹, Pat Levitt¹, Kathie Eagleson¹, Sarah Kennedy¹, and Ying Wang¹

¹ Department of Neurobiology and Anatomy, Allegheny University of the Health Sciences, Philadelphia, Pennsylvania 19129, and ² The Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104

A survival-promoting peptide has been purified from medium conditioned by Y79 human retinoblastoma cells and a mouse hippocampal cell line (HN 33.1) exposed to H₂O₂. A 30 residue synthetic peptide was made on the basis of N-terminal sequences obtained during purification, and it was found to exhibit gel mobility and staining properties similar to the purified molecules. The peptide maintains cells and their processes in vitro for the HN 33.1 cell line treated with H₂O₂, and in vivo for cortical neurons after lesions of the cerebral cortex. It has weak homology with a fragment of a putative bacterial antigen and, like that molecule, binds IgG. The peptide also contains a motif reminiscent of a critical sequence in the catalytic region of calcineurin-type phosphatases; surprisingly, like several members of this family, the peptide catalyzes the hydrolysis of para-nitrophenylphosphate in the presence of Mn²⁺. Application of the peptide to one side of bilateral cerebral cortex lesions centered on area 2 in rats results in an increase in IgG immunoreactivity in the vicinity of the lesions 7 d after surgery. Microglia immunopositive for IgG and ED-1 are, however, dramatically reduced around the lesions in the treated hemisphere. Furthermore, pyramidal neurons that would normally shrink, die, or disintegrate were maintained, as determined by MAP2 immunocytochemistry and Nissl staining. These survival effects were often found in both hemispheres. The results suggest that this peptide operates by diffusion to regulate the immune response and thereby rescue neurons that would usually degenerate after cortical lesions. The phosphatase activity of this molecule also suggests the potential for direct neuron survival-promoting effects.

Key words: peptide; neuron survival; immune evasion; microglia; IgG; phosphatase

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Copyright © 1998 Society for Neuroscience  0270-6474/98/18187047-14$05.00/0

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