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Neuropeptide Y gene expression in immortalized rat hippocampal and pheochromocytoma-12 cell lines.

机译:神经肽Y基因在永生大鼠海马和嗜铬细胞瘤12细胞系中的表达。

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Employing clonal cell lines derived from rat embryonic hippocampal cells, we detected neuropeptide Y (NPY) mRNA in three progenitor subcloned cell lines. These cell lines upon differentiation express markers indicative of commitment to either neuronal (H19-7; NF +, GFAP -), glial (H19-5; GFAP +, NF -), or bipotential (H583-5; NF +, GFAP + ) lineages. Induction of differentiation was associated with the persistence of the NPY mRNA, however, in the differentiated H19-7 cells a 20-fold increase in NPY mRNA levels was observed (P<0.05). NPY immunoreactivity was observed only in cells with a differentiated neuronal phenotype. The cellular radioimmunoassayable NPY peptide levels increased twelve-fold without a change in extracellular NPY peptide levels by multi-factorially induced neuronal or glial cell differentiation. The differentiated H19-5 cells expressed lower levels of NPY that could not be immunocytochemically detected. The peripheral sympathetic PC-12 neuronal cells examined in the undifferentiated and nerve growth factor-driven differentiated states expressed NPY only upon differentiation. We conclude that NPY is expressed by the cultured undifferentiated and differentiated rat hippocampal clonal cell lines, while the peripheral sympathetic PC-12 neuronal cell line only expresses the NPY gene upon differentiation. These immortalized embryonic neural cell line(s) will provide a hippocampal cell line(s) to conduct future in-vitro investigations targeted at determining the cellular and molecular mechanisms governing NPY gene expression.
机译:利用源自大鼠胚胎海马细胞的克隆细胞系,我们在三种祖细胞亚克隆细胞系中检测到神经肽Y(NPY)mRNA。这些分化后的细胞系表达指示对神经元(H19-7; NF +,GFAP-),神经胶质(H19-5; GFAP +,NF-)或双能(H583-5; NF +,GFAP + )血统。分化的诱导与NPY mRNA的持续存在有关,但是,在分化的H19-7细胞中,观察到NPY mRNA水平增加了20倍(P <0.05)。仅在具有分化的神经元表型的细胞中观察到NPY免疫反应性。通过多因素诱导的神经元或神经胶质细胞分化,细胞放射免疫可测定的NPY肽水平增加了十二倍,而胞外NPY肽水平没有变化。分化的H19-5细胞表达的NPY水平较低,无法通过免疫细胞化学方法检测到。在未分化和神经生长因子驱动的分化状态下检查的外周交感性PC-12神经元细胞仅在分化后才表达NPY。我们得出结论,NPY由培养的未分化和分化的大鼠海马克隆细胞系表达,而周围交感的PC-12神经元细胞系仅在分化时表达NPY基因。这些永生化的胚胎神经细胞系将提供海马细胞系,以进行未来的体外研究,旨在确定控制NPY基因表达的细胞和分子机制。

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