| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
We and others previously reported that soluble stem cell factor (sSCF) and c-KIT interaction plays an important role in the development of melanocytes from neural crest cells (NCCs). It has also been shown that endothelin-3 (ET-3) promotes not only proliferation but also differentiation of NCCs. In this study, to determine the effects of ET-3 in combination with sSCF on the proliferation and differentiation of cultured NCCs into melanocytes, we cultured NCCs from wild type and 'Steel' mutant mice with ET-3, sSCF or ET-3+sSCF for 3-15 days. After immunohistochemical and DOPA staining, we counted the number of c-KIT(+) and DOPA(+) cells. In the wild type, the addition of ET-3 increased the number of c-KIT(+) and DOPA(+) cells in a dose- and time- dependent manner, while the addition of sSCF mainly increased the number of c-KIT(+) cells. Pigmented melanocytes were detected with ET-3 but not with sSCF.The combination with sSCF and ET-s showed a marked increase of c-KIT(+), DOPA(+) cells and pigmented melanocytes. In 'Steel' mutant mice, all those three cells did not appear in NCCs cultured with ET-3, although pigmented melanocytes were detected when SCF and ET-3 were both added to 'Steel' NCCs. These results suggest that there are intrinsic SCF sources in the wild type NCC explants, and that the survival of c-KIT(+) melanocyte precursers supported by SCF is essential to the acceleration of melanization by ET-3.
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