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The Journal of Neuroscience, July 1, 2001, 21(13):4772-4781

Intact Aggrecan and Fragments Generated by Both Aggrecanse and Metalloproteinase-Like Activities Are Present in the Developing and Adult Rat Spinal Cord and Their Relative Abundance Is Altered by Injury

Michele L. Lemons^{1, 3}, John D. Sandy⁴, Douglas K. Anderson^{1, 2, 3, 5}, and Dena R. Howland^{1, 3}

Departments of ¹ Neuroscience and ² Neurological Surgery, University of Florida College of Medicine, Gainesville, Florida 32610-0244, ³ Evelyn F. and William L. McKnight Brain Institute of The University of Florida, Gainesville, Florida 32611, ⁴ Shriners Hospital for Crippled Children and College of Medicine, University of South Florida, Tampa, Florida 33612, and ⁵ Malcom Randall Veterans Affairs Medical Center, Gainesville, Florida 32608

Aggrecan is a large proteoglycan (PG) that has been grouped with different PG families on the basis of its physical characteristics. These families include the chondroitin sulfate PGs, which appear to inhibit the migration of cells and axons during development. Although aggrecan has been studied primarily in cartilage, in the present study, tissue samples from developing, mature, and injured-adult rat spinal cords were used to determine whether aggrecan is present in the mammalian spinal cord. By the use of Western blot analysis, tissues were probed with aggrecan-specific antibodies (ATEGQV, TYKHRL, and LEC-7) and aggrecan-specific neoepitope antibodies (NITEGE, FVDIPEN, and TFKEEE) to identify full-length aggrecan and several fragments. Unlike many other aggrecan gene family members, aggrecan species were similar in embryonic day 14, postnatal day 1, and adult spinal cords. Spinal cord injury caused significant decreases in aggrecan. Partial recovery in some aggrecan species was evident by 2 weeks after injury. The presence of specific aggrecan neoepitopes suggested that aggrecan is cleaved in the spinal cord by both a disintegrin and metalloproteinase thrombospondin (also known as aggrecanase) and metalloproteinase-like activities. Many aggrecan species found in the spinal cord were similar to species in cartilage. Additional antibodies were used to identify two other aggrecan gene family members, neurocan and brevican, in the adult spinal cord. These studies present novel information on the aggrecan core protein species and enzymes involved in aggrecan cleavage in vivo in the rat spinal cord throughout development and after injury. They also provide the basis for investigating the function of aggrecan in the spinal cord.

Key words: spinal cord; spinal cord injury; development; aggrecan; aggregating proteoglycan; chondroitin sulfate proteoglycan; metalloproteinase; aggrecanase; neurocan; brevican

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Copyright © 2001 Society for Neuroscience  0270-6474/01/21134772-10$05.00/0

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