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Volume 16, Number 24, Issue of December 15, 1996 pp. 8005-8018
Copyright ©1996 Society for Neuroscience

Chondroitin Sulfate Proteoglycan and Tenascin in the Wounded Adult Mouse Neostriatum In Vitro: Dopamine Neuron Attachment and Process Outgrowth

Received Aug. 14, 1996; revised Oct. 1, 1996; accepted Oct. 3, 1996.

Monte A. Gates, Helen Fillmore, and Dennis A. Steindler

Department of Anatomy and Neurobiology, University of Tennessee, Memphis, Tennessee 38163

Extracellular matrix (ECM) molecules, including chondroitin-4 or chondroitin-6 sulfate proteoglycans (CSPGs) and tenascin, are upregulated in and around wounds and transplants to the adult CNS. In the present study, striatal wounds from adult mice were used in a novel in vitro paradigm to assess the effects of these wound-associated molecules on embryonic dopamine cell attachment and neurite outgrowth. Light and electron microscopic immunocytochemistry studies have shown that astroglial scar constituents persist in cultured explants for at least 1 week in vitro, and despite the loss of neurons from adult striatal explants, there is a retention of certain structural features suggesting that the wound explant-neuron coplant is a viable model for analysis of graft-scar interactions. Explants from the wounded striatum taken at different times after a penetrating injury in vivo were used as substrates for embryonic ventral mesencephalon neurons that were plated on their surfaces. Dopamine cell attachment is increased significantly in relation to the expression of both CSPG and tenascin. The increase in neuronal attachment in this paradigm, however, is accompanied by a postlesion survival time-dependent significant decrease in neuritic growth from these cells. In vitro ECM antibody treatment suggests that CSPG may be responsible for heightened dopamine cell attachment and that tenascin simultaneously may support cell attachment while inhibiting neurite growth. The present study offers a new approach for the in vitro analysis of cell and molecular interactions after brain injury and brain grafting, in essence acting as a nigrostriatal transplant-in-a-dish.

Key words: extracellular matrix; explant culture; brain injury; dopamine neurons; cell-substrate interactions; neurite growth

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