Nabeka Hiroaki, Uematsu Keigo, Takechi Hiroko, Shimokawa Tetsuya, Yamamiya Kimiko, Li Cheng, Doihara Takuya, Saito Shouichiro, Kobayashi Naoto, Matsuda Seiji
Department of Anatomy and Embryology, Ehime University Graduate School of Medicine, Toon, Ehime, Japan.
Laboratory of Veterinary Anatomy, Faculty of Applied Biological Sciences, Gifu University, Yanagido, Gifu, Japan.
PLoS One. 2014 Dec 2;9(12):e110534. doi: 10.1371/journal.pone.0110534. eCollection 2014.
Because excessive glutamate release is believed to play a pivotal role in numerous neuropathological disorders, such as ischemia or seizure, we aimed to investigate whether intrinsic prosaposin (PS), a neuroprotective factor when supplied exogenously in vivo or in vitro, is up-regulated after the excitotoxicity induced by kainic acid (KA), a glutamate analog. In the present study, PS immunoreactivity and its mRNA expression in the hippocampal and cortical neurons showed significant increases on day 3 after KA injection, and high PS levels were maintained even after 3 weeks. The increase in PS, but not saposins, detected by immunoblot analysis suggests that the increase in PS-like immunoreactivity after KA injection was not due to an increase in saposins as lysosomal enzymes after neuronal damage, but rather to an increase in PS as a neurotrophic factor to improve neuronal survival. Furthermore, several neurons with slender nuclei inside/outside of the pyramidal layer showed more intense PS mRNA expression than other pyramidal neurons. Based on the results from double immunostaining using anti-PS and anti-GABA antibodies, these neurons were shown to be GABAergic interneurons in the extra- and intra-pyramidal layers. In the cerebral cortex, several large neurons in the V layer showed very intense PS mRNA expression 3 days after KA injection. The choroid plexus showed intense PS mRNA expression even in the normal rat, and the intensity increased significantly after KA injection. The present study indicates that inhibitory interneurons as well as stimulated hippocampal pyramidal and cortical neurons synthesize PS for neuronal survival, and the choroid plexus is highly activated to synthesize PS, which may prevent neurons from excitotoxic neuronal damage. To the best of our knowledge, this is the first study that demonstrates axonal transport and increased production of neurotrophic factor PS after KA injection.
由于过量谷氨酸释放被认为在多种神经病理疾病中起关键作用,如局部缺血或癫痫发作,我们旨在研究内源性 prosaposin(PS),一种在体内或体外外源性供应时具有神经保护作用的因子,在谷氨酸类似物 kainic acid(KA)诱导的兴奋性毒性后是否会上调。在本研究中,KA 注射后第 3 天,海马和皮质神经元中的 PS 免疫反应性及其 mRNA 表达显著增加,即使在 3 周后仍维持高 PS 水平。免疫印迹分析检测到的 PS 增加而非 saposins 的增加表明,KA 注射后 PS 样免疫反应性的增加不是由于神经元损伤后作为溶酶体酶的 saposins 增加,而是由于作为改善神经元存活的神经营养因子的 PS 增加。此外,锥体细胞层内外一些核细长的神经元比其他锥体神经元显示出更强的 PS mRNA 表达。基于使用抗 PS 和抗 GABA 抗体的双重免疫染色结果,这些神经元被证明是锥体细胞层外和层内的 GABA 能中间神经元。在大脑皮层,KA 注射后 3 天,V 层的几个大神经元显示出非常强烈的 PS mRNA 表达。脉络丛即使在正常大鼠中也显示出强烈的 PS mRNA 表达,KA 注射后强度显著增加。本研究表明,抑制性中间神经元以及受刺激的海马锥体神经元和皮质神经元合成 PS 以促进神经元存活,并且脉络丛被高度激活以合成 PS,这可能防止神经元受到兴奋性毒性神经元损伤。据我们所知,这是第一项证明 KA 注射后神经营养因子 PS 的轴突运输和产量增加的研究。