Nakanishi T, Takahashi K, Aoki C, Nishikawa K, Hattori T, Taniguchi S
Department of Biochemistry, Okayama University Dental School, Japan.
Biochem Biophys Res Commun. 1994 Feb 15;198(3):891-7. doi: 10.1006/bbrc.1994.1127.
With cultured osteoblastic cells, clone MC3T3-E1 derived from newborn mouse calvaria, the mRNAs encoding three representative neurotrophins, namely, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3), were proved in estimation by the polymerase chain reaction (PCR) method. The increase and then subsequent decrease of all three mRNAs within the proliferation phase were followed by their gradual increase in the differentiation phase, with a tendency of enhancement by exogenous TGF-beta as was particularly evident in the case of NGF. These findings were further substantiated by identification of NGF-like neutrophils in the conditioned medium of the osteoblastic cells by an immunoblotting and neurite extension assay.
利用从小鼠新生颅骨分离培养的成骨细胞克隆MC3T3-E1,通过聚合酶链反应(PCR)方法证实了编码三种代表性神经营养因子的mRNA,即神经生长因子(NGF)、脑源性神经营养因子(BDNF)和神经营养因子-3(NT-3)。在增殖期,所有三种mRNA先增加后减少,随后在分化期逐渐增加,外源性转化生长因子-β(TGF-β)有增强作用,在NGF的情况下尤为明显。通过免疫印迹和神经突延伸试验在成骨细胞条件培养基中鉴定出NGF样中性粒细胞,进一步证实了这些发现。