626 Characterization of activated microglia in axotomized facial nucleus of adult rat

s91

625

Establishment

of stage-specific pure oligodendrocyte

cultures and oligodendrocyte

lineage

Dept. of Cell Biology’, Neurology Tokyo Metropolitan Inst. of Gerontology, Itabashi, Tokyo 173, Japan’, Dept. of Biophysics and Lifescience, Univ. of Tokyo at Komaba, Meguro, Tokyo 153 Japan3 Y. Sakurai’, D. Nishimura3, S. Kawato3, T. Hamada2, H. Yamanouchi ‘2, M. Takahashil’! C. Seiwa’, H. Asoul We report mass-production technique that allow one to develop and maintain cultures stage-specific pureoligo-dendrocytes from embryonic rat brain. After four weeks cultures, they consist of more than 99% oligodendrocytes and fewer than 1% non-neuronal cells. When the oligodendrocyte lineage-specific glyco-lipid antibodies A2B5, GC, 04 and 01 were used to identify the differentiation of progenitor cells, we found that pure-oligodendrocytes were expressed 04, which is labeled mitotic oligodendrocyte progenitor, postmitotic, differentiated oligodendrocytes. They also acquired the ability to bind 01, but they were not expressed GC and MBP on their surface. For their survival, the cells requires the addition of typel-astrocytes as a coculture. Our results suggest that the stage-specific differentiation of oligodendrocytes into mature type does occur in a restricted manner, assist with typel-astrocytes in cultures.

626 Department 187, Japan

CHARACTERIZATION OF ACTIVATED MICROGLIA IN AXOTOMIZED FACIAL NUCLEUS OF ADULT RAT of Neurochemistry, National Institute of Neuroscience, 4-l-l Ogawa-higashi, Kodaira, Tokyo

KAZUYUKI NAKAJIMA, MASAHIRO ISHIKAWA, SHIZUYO HONDA, SHINICHI KOHSAKA To study the function of activated microglia observed in the lesioned site of adult brain, the activated microglia were isolated from axotomized facial nuclei of adult rats by explant culture method. These cells were stained by ED-l monoclonal antibody, and showed strong phagocytic activity, Isolectin B4-binding activity and acetylated low density lipoprotein incorporation activity. Judging from these properties, the purity of the microglia was estimated to be almost 100%. Distinct from new born rat brain-derived microglia, these cells had an ability to proliferate by themselves without co-culture with other type of cells. The analysis of specific receptor for colonystimulating factor (CSF) revealed the expression of c-Fms on the cultured microglia. These results suggest that the microglia from axotomized facial nuclei of adult rats might proliferate in response to macrophage-CSF.

627 Department MASATOSHI

Induction of apoptosis by protein kinase inhibitors in the cells of Drosophila neuronal cell line following treatment with 20-hydroxyecdysone. of pharmacology, Nippon Medical School, Tokyo 113, Japan NAGANO,

KUMIKO

UI-TEI,

HIDENORI

SUZUKI,

YUHEI

MIYATA

We have induced apoptosis by H-7, a broad spectrum protein kinase inhibitor, in the neuronal cells of MLDmBG2-c2 cell line derived from Drosophila larval CNS. However, it was revealed that the apoptosis is due to inhibition of H-7 sensitive substance(s) other than the protein kinases known to be affected by H-7, because specific inhibitors to the kinases do not induce apoptosis. It is possible that the effects of kinase inhibitors on the cells is due to the state of cells, proliferating. 20-hydroxyecdysone (20-HE), an arthropod molting hormone, plays a major role for metamorphosis during Drosophila development, and is known to arrest ceil cycle at G2 phase in Drosophila Kc cell line. Treatment of BG2-c2 cells with 20-HE inhibited cell proliferation and induced morphological changes. In contrast to the untreated cells, some specific kinase inhibitors as well as H-7 induced apoptosis in the 20-HE treated cells.