Ishikawa T, Kawai C, Sano M, Minatogawa Y
Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan.
Exp Cell Res. 2001 Jun 10;266(2):260-9. doi: 10.1006/excr.2001.5226.
Localization and movement of peroxisomes have been investigated in neurites of a subline of PC12 pheochromocytoma cells (PC12D cells). The cells were transfected with a construct encoding the green fluorescent protein and bearing the C-terminal peroxisomal targeting signal 1 SKL motif (-Ser-Lys-Leu-COOH). Peroxisomes were detected as green punctate fluorescent signals. Many peroxisomes were observed in neurites of PC12D cells, especially in neural terminal-like structures, growth cones, varicosities, and branch points. Growth cones containing many peroxisomes were active, since they extended several long filopodias. Existence of peroxisomes in growth cones and neuronal terminal-like structures suggests that peroxisomes might have some role in neuronal extension and nerve terminal functioning. Peroxisomal motility was analyzed by time-lapse imaging using a fluorescence microscope at 25 degrees C. Peroxisomes were transported bidirectionally in neurites, i.e., through anterograde and retrograde transport. This result suggests that peroxisomes move to growth cones and neural terminals from the PC12D cell body, play some role in these parts, and go back to cell body.
在PC12嗜铬细胞瘤细胞亚系(PC12D细胞)的神经突中,对过氧化物酶体的定位和移动进行了研究。用编码绿色荧光蛋白并带有C末端过氧化物酶体靶向信号1 SKL基序(-Ser-Lys-Leu-COOH)的构建体转染细胞。过氧化物酶体被检测为绿色点状荧光信号。在PC12D细胞的神经突中观察到许多过氧化物酶体,尤其是在神经末梢样结构、生长锥、膨体和分支点中。含有许多过氧化物酶体的生长锥是活跃的,因为它们伸出了几条长长的丝状伪足。生长锥和神经末梢样结构中存在过氧化物酶体表明,过氧化物酶体可能在神经元延伸和神经末梢功能中发挥某些作用。在25℃下使用荧光显微镜通过延时成像分析过氧化物酶体的运动性。过氧化物酶体在神经突中双向运输,即通过顺行和逆行运输。这一结果表明,过氧化物酶体从PC12D细胞体移动到生长锥和神经末梢,在这些部位发挥某些作用,然后回到细胞体。