Screening for neutrophic factors and inhibitors of NGF using PC-12 or SH-SY5Y cells.

Outlines:

Neurotrophic factors are thought to be essential for survival and functional maintenance of nerve cells in the central and the peripheral nervous system. The compounds which regulate the synthesis and secretion of neurotrophic factors might be a good candidate for pharmaceutical agents of various neurodegenerative diseases involving dementia. From this point of view, we are screening for neuritogenic compounds. For the bioassay to obtain the neuritogenic compound , human neuroblastoma cell line, SH-SY5Y or rat pheochromocytoma cell line, PC-12 was used as an indicator.

(A)

(B)

(C)

Morphological changes of SH-SY5Y cells treated with epolactaene or dibutyryl cAMP.
The cells were cultured at 37 in 5 % CO2 humidified atmosphere in Basal Medium Eagle (BME) medium containing 10 % fetal bovine serum on plastic culture dishes. Photograph were obtained after 2 days cultivation with/without epolactaene or dibutyryl cAMP. (A), control; (B), with 10 ug/ml of epolactaene; (C), with 1 mM of dibutyryl cAMP. (Ref:2)

Day 1

Day 3

Control
NGF
MT-19
MT-20
Differentiation of normal PC12 cells induced by NGF or gamma-lactam-related compounds.
Cells were plated on poly-L-lysine-coated 24-well dishes at the density of 1000 cells per well. After 3 to 16 h, media were changed to fresh media containing 0.5 % FBS and NGF (100 ng/ml), MT-19 (1 ug/ml), or MT-20 (1 ug/ml). Cells were photographed after 1 nad 3 days of treatment. (Ref:1)

References:

  1. H. Kakeya, I. Takahashi, G. Okada, K. Isono, H. Osada.
    Epolactaene, a novel neuritogenic compound in human neuroblastoma cells, produced by a marine fungus.
    J. Antibiotics, 48, 733-735 (1995).

  2. M. Matsumoto, T. Sudo, M. Maruyama, H. Osada, M. Tsujimoto.
    Activation of p38 mitogen-activated protein kinase is crucial in osteoclastogenesis induced by tumor necrosis factor.
    FEBS Lett., 486, 23-28 (2000).

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