Sano M, Iwanaga M, Fujisawa H, Nagahama M, Yamazaki Y
Institute for Developmental Research, Aichi Prefectural Colony, Japan.
Brain Res. 1994 Mar 7;639(1):115-24. doi: 10.1016/0006-8993(94)91771-x.
Staurosporine, a potent inhibitor of protein kinases, caused the rapid outgrowth of neurites from cultured dorsal root ganglia of chick embryos and from PC12D cells, a subline of PC12 cells. Treatment of dorsal root ganglia with 1 to 20 nM staurosporine resulted in the extensive outgrowth of neurites that were indistinguishable from those induced by NGF, as assessed by phase-contrast microscopy, electron microscopy and cytochemical staining of actin and tubulin. However, neurites generated from the ganglia in response to the higher concentrations of staurosporine (40-100 nM) seemed to have different characteristics, possibly as a result of the inhibition of cell migration from ganglia. The sequential changes in morphology of PC12D cells in response to staurosporine and to NGF were revealed by staining of actin. Ruffling membranes emerged at the margins of PC12D cells within 4 min after the addition of staurosporine or of NGF. From 10 min to 24 h after the addition of either compound, the ruffles were transformed into several projections that became growing neurites. The formation of ruffles and the outgrowth of neurites were both apparent at a concentration of staurosporine of 10 nM. The neurites that emerged from PC12D cells in response to staurosporine and in response to NGF were indistinguishable under the phase-contrast microscope and after staining of actin and tubulin. However, staurosporine never promoted survival of PC12D cells in serum-free conditions as that promoted by NGF. The observations indicate that staurosporine at nanomolar concentrations may reproduce the neurogenic changes that induced by NGF in primed neuronal cells, although it can not mimic the action of NGF that supports survival of neurons.
星形孢菌素是一种有效的蛋白激酶抑制剂，可使培养的鸡胚背根神经节以及PC12细胞的一个亚系PC12D细胞快速长出神经突。用1至20 nM的星形孢菌素处理背根神经节，会导致神经突大量长出，通过相差显微镜、电子显微镜以及肌动蛋白和微管蛋白的细胞化学染色评估，这些神经突与神经生长因子（NGF）诱导产生的神经突无法区分。然而，背根神经节对较高浓度（40 - 100 nM）星形孢菌素产生反应而生成的神经突似乎具有不同特征，这可能是由于神经节细胞迁移受到抑制所致。通过肌动蛋白染色揭示了PC12D细胞对星形孢菌素和NGF反应时形态的连续变化。添加星形孢菌素或NGF后4分钟内，PC12D细胞边缘出现膜皱襞。添加任何一种化合物后10分钟至24小时，这些皱襞转变为几个突起，进而成为生长中的神经突。在10 nM的星形孢菌素浓度下，膜皱襞的形成和神经突的长出都很明显。在相差显微镜下以及对肌动蛋白和微管蛋白进行染色后，由PC12D细胞对星形孢菌素和NGF产生反应而长出的神经突无法区分。然而，在无血清条件下，星形孢菌素从未像NGF那样促进PC12D细胞的存活。这些观察结果表明，纳摩尔浓度的星形孢菌素可能重现NGF在已致敏神经元细胞中诱导的神经源性变化，尽管它无法模拟NGF支持神经元存活的作用。
