Ando Yoshio, Liang Yideng, Ishigaki Shinsuke, Niwa Jun-ichi, Jiang Yuemei, Kobayashi Yasushi, Yamamoto Masahiko, Doyu Manabu, Sobue Gen
Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
Neurochem Res. 2003 Jun;28(6):839-46. doi: 10.1023/a:1023258923002.
Amyotrophic lateral sclerosis is characterized by selective motor neuron degeneration. An apoptotic pathway is thought to be involved. It is difficult, however, to analyze the molecular pathogenic mechanism in single motor neurons because of complexity in the neural tissue, which consists of multiple lineages of cells neighboring motor neurons. We quantified the caspase-1 and -3 mRNA in single motor neurons and neighboring glial cells isolated from the spinal ventral horn of mutant SOD1 transgenic (Tg) mice and littermates. Motor neurons and neighboring glial cells were isolated from spinal sections by laser microdissection, and the mRNAs were quantified by RT-PCR. In the Tg mice, caspase-1 mRNA was first upregulated in motor neurons and second in glial cells. The caspase-3 mRNA was increased in motor neurons following the caspase-1 mRNA. These results indicated that caspase-1 and -3 mRNAs are differentially upregulated in motor neurons and glial cells of the Tg mice, and that mRNAs in isolated cells can be accurately assessed using our procedures.
肌萎缩侧索硬化症的特征是选择性运动神经元变性。据认为,一条凋亡途径与之相关。然而,由于神经组织的复杂性,其中包含与运动神经元相邻的多种细胞谱系,因此很难分析单个运动神经元中的分子致病机制。我们对从突变型超氧化物歧化酶1转基因(Tg)小鼠和同窝小鼠的脊髓腹角分离出的单个运动神经元和相邻神经胶质细胞中的半胱天冬酶-1和-3信使核糖核酸进行了定量分析。通过激光显微切割从脊髓切片中分离出运动神经元和相邻神经胶质细胞,并通过逆转录聚合酶链反应对信使核糖核酸进行定量分析。在Tg小鼠中,半胱天冬酶-1信使核糖核酸首先在运动神经元中上调,其次在神经胶质细胞中上调。半胱天冬酶-3信使核糖核酸在半胱天冬酶-1信使核糖核酸之后在运动神经元中增加。这些结果表明,半胱天冬酶-1和-3信使核糖核酸在Tg小鼠的运动神经元和神经胶质细胞中差异上调,并且使用我们的方法可以准确评估分离细胞中的信使核糖核酸。