1990 Fiscal Year Final Research Report Summary
Study of Cultured Glial Cells
| Project/Area Number |
01570445
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurology
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| Research Institution | Fukui Medical School |
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Principal Investigator |
HIRAYAMA Mikio Fukui Med. Sch. Dept. Med. Sci., Assistant Professor, 医学部, 講師 (30181192)
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| Co-Investigator(Kenkyū-buntansha) |
KUBOTA Toshihiko Fukui Med. Sch. Dept. Med. Sci., Associate Professor, 医学部, 助教授 (70092781)
YOKOCHI Takashi Fukui Med. Sch. Dept. Med. Sci., Associate Professor, 医学部, 助教授 (20126915)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Oligodendrocyte / Astrocyte / Dibutyryl cyclic AMP / Ganglioside / Trapidil / PDGF / Lysosomal enzyme |
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| Research Abstract |
1. Lysosomal enzyme activity of cultured bovine oligodendrocytes was assayed. At 4DIV, alpha-mannosidase, alpha-fucosidase, beta-galactosidase activity was higher in 2x10^6 cells than in lx10^6 cells plated. Lysosomal activity become higher at 6-8DIV than 4DIV.
2. The addition of 1mM of cyclic AMP analogs on 100muM of forskolin to rat oligodendrocytes in vitro enhanced immunofluorescence intensity of anti-galactocerebroside and anti-myelin basic protein antibody to oligodendrocytes assayed by microfluorocytometry, which indicates the increased synthesis of these antigens. Oligodendrocytes treated with cyclic AMP analogs showed longer processes and formed a network among processes. Cyclic AMP analogs enhance the function of oligodendrocytes in vitro and thus might be useful agents for enhancing remyelination.
3. We examined the factors which influence the differentiation of the oligodendrocyte-type-2 astrocyte (0-2A) progenitor cells in vitro. At 5 to 7 day in vitro culture from newborn r
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at brain, 1mM dibutyryl cyclic AMP. 10-500mug/ml mixed bovine ganglioside, 10-800mug/ml trapidil were added for 3 to 7 days. Dibutyryl cyclic AMP increased the number of galactocerebroside(+) oligodendrocytes, whereas it decreased the number of 0-2A progenitor cells. This suggested the enhanced differentiation of 0-2A progenitor cells to oligodendrocytes. Ganglioside had similar effects with less degree. Trapidil, antagonist of PDGF was cytotoxic to 0-2A progenitor cells and oligodendrocytes at higher concentration. Fibrous astrocytes were predominant cells in trapidil-treated cultures, whereas protoplasmic astrocytes were predominant cells in control cultures.
4. To study the factors responsible for remyelination after demyelination, we examined whether oligodendrocyte progenitor cells exist in adult bovine brains and examined the effect of dibutyryl cyclic AMP, ganglioside and trapidil on oligodendrocytes in vitro. Oligodendrocyte-rich fractions were iso
lated by Percoll density gradient from adult bovine white matter. We found that there was low percentage (0.1-0.4%) of 0-2A progenitor cells. 100mug/ml ganglioside, 500mug/ml trapidil inhibited the elongation of processes and trapidil exerted its cytotoxicity. 1mM of dibutyryl cyclic AMP had no effect on morphology of oligodendrocytes.
5. Transplantation experiment of mature bovine oligodendrocytes to shiverer mutant mouse is under investigation and PDGF receptor of oligodendrocytes is also to be studied. Less
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