DOWN-REGULATION OF AMYLOID PRECURSOR PROTEIN INHIBITS NEURITE OUTGROWTH IN-VITRO
DOWN-REGULATION OF AMYLOID PRECURSOR PROTEIN INHIBITS NEURITE OUTGROWTH IN-VITRO
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DOI：
10.1083/jcb.128.5.919
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发表时间：
1995-03-01
影响因子：
7.8
通讯作者：
PROCHIANTZ, A
中科院分区：
文献类型：
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作者：
ALLINQUANT, B;HANTRAYE, P;PROCHIANTZ, A
淀粉样蛋白前体蛋白（APP）是一种在多种细胞类型中表达的跨膜蛋白。在神经系统中，APP由胶质细胞和神经元细胞表达，多项证据表明它在正常和病理现象中发挥作用。为了解决APP在正常发育神经元中的实际功能问题，我们进行了一项旨在利用反义寡核苷酸阻断APP表达的研究。寡核苷酸内化是通过将它们连接到通过生物膜易位的载体肽来实现的。这种原始技术非常高效，可以直接进入细胞质和细胞核，使我们能够处理细胞外寡核苷酸浓度在40到200 nM之间的情况。与翻译起源重叠的反义寡核苷酸内化导致APP新合成显著但短暂的减少，这是单独使用载体肽或义寡核苷酸所没有观察到的。尽管是短暂的，APP新合成的减少足以引起体外发育的胚胎皮质神经元轴突和树突生长的明显减少。后者的减少不伴有细胞体扩散的变化。在培养开始时，单次暴露于偶联反义寡核苷酸足以在6、18和24小时后产生显著的形态学影响，但到42小时时，没有其他显著的形态学变化。本报告表明，淀粉样前体蛋白在原代培养皮层神经元的形态分化中起重要作用。
The amyloid precursor protein (APP) is a transmembrane protein expressed in several cell types. In the nervous system, APP is expressed by glial and neuronal cells, and several lines of evidence suggest that it plays a role in normal and pathological phenomena. To address the question of the actual function of APP in normal developing neurons, we undertook a study aimed at blocking APP expression using antisense oligonucleotides. Oligonucleotide internalization was achieved by linking them to a vector peptide that translocates through biological membranes. This original technique, which is very efficient and gives direct access to the cell cytosol and nucleus, allowed us to work with extracellular oligonucleotide concentrations between 40 and 200 nM. Internalization of antisense oligonucleotides overlapping the origin of translation resulted in a marked but transient decrease in APP neosynthesis that was not observed with the vector peptide alone, or with sense oligonucleotides. Although transient, the decrease in APP neosynthesis was sufficient to provoke a distinct decrease in axon and dendrite outgrowth by embryonic cortical neurons developing in vitro. The latter decrease was not accompanied by changes in the spreading of the cell bodies. A single exposure to coupled antisense oligonucleotides at the onset of the culture was sufficient to produce significant morphological effects 6, 18, and 24 h later, but by 42 h, there were no remaining significant morphologic changes. This report thus demonstrates that amyloid precursor protein plays an important function in the morphological differentiation of cortical neurons in primary culture.