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Inhibition of proteoglycan synthesis affects neuronal outgrowth and astrocytic migration in organotypic cultures of fetal ventral mesencephalon
Author(s) -
Berglöf Elisabet,
Plantman Stefan,
Johansson Saga,
Strömberg Ingrid
Publication year - 2008
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.21465
Subject(s) - midbrain , dopamine , chondroitin sulfate proteoglycan , microbiology and biotechnology , proteoglycan , neuroscience , tyrosine hydroxylase , nerve growth factor , biology , axoplasmic transport , chemistry , extracellular matrix , central nervous system , biochemistry , receptor
Abstract Grafting fetal ventral mesencephalon has been utilized to alleviate the symptoms of Parkinson's disease. One obstacle in using this approach is the limited outgrowth from the transplanted dopamine neurons. Thus, it is important to evaluate factors that promote outgrowth from fetal dopamine neurons. Proteoglycans (PGs) are extracellular matrix molecules that modulate neuritic growth. This study was performed to evaluate the role of PGs in dopamine nerve fiber formation in organotypic slice cultures of fetal ventral mesencephalon. Cultures were treated with the PG synthesis inhibitor methyl‐umbelliferyl‐β‐ D ‐xyloside (β‐xyloside) and analyzed using antibodies against tyrosine hydroxylase (TH) to visualize dopamine neurons, S100β to visualize astrocytes, and neurocan to detect PGs. Two growth patterns of TH‐positive outgrowth were observed: nerve fibers formed in the presence of astrocytes and nerve fibers formed in the absence of astrocytes. Treatment with β‐xyloside significantly reduced the distance of glial‐associated TH‐positive nerve fiber outgrowth but did not affect the length of the non‐glial‐associated nerve fibers. The addition of β‐xyloside shifted the nerve fiber growth pattern from being mostly glial‐guided to being non‐glial‐associated, whereas the total amount of TH protein was not affected. Further, astrocytic migration and proliferation were impaired after β‐xyloside treatment, and levels of non‐intact PG increased. β‐Xyloside treatment changed the distribution of neurocan in astrocytes, from being localized in vesicles to being diffusely immunoreactive in the processes. To conclude, inhibition of PG synthesis affects glial‐associated TH‐positive nerve fiber formation in ventral mesencephalic cultures, which might be an indirect effect of impaired astrocytic migration. © 2007 Wiley‐Liss, Inc.