Bergold P J, Casaccia-Bonnefil P, Zeng X L, Federoff H J
Department of Pharmacology, State University of New York-Health Science Center, Brooklyn 11203.
Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6165-9. doi: 10.1073/pnas.90.13.6165.
Patients with severe temporal lobe epilepsy lose neurons within the CA3 and hilar regions of the hippocampus. Loss of CA3 and hilar neurons was also induced by transducing organotypic hippocampal slice cultures with a replication-defective herpes simplex virus (HSV) vector expressing the GluR6 kainate subtype of the glutamate receptor (HSVGluR6). In transduced fibroblasts, HSVGluR6 expressed a M(r) 115,000 protein that reacted with anti-GluR6 serum. After exposure of fibroblast to HSVGluR6, a kainate-dependent toxicity appeared in cells that were previously resistant to kainate. Microapplication of nanoliter amounts of recombinant HSV stocks into organotypic hippocampal slice cultures resulted in localized transduction and gene transfer at the site of microapplication. Microapplication of 100 HSVGluR6 virions into CA3 stratum pyramidale induced a large loss of CA3 pyramidal cells and hilar neurons, despite the small number of transduced neurons. This effect was not seen when 100 virions of HSVGluR6 were microapplied to CA1 stratum pyramidale. Tetrodotoxin or N-methyl-D-aspartate receptor antagonists inhibited the large loss of CA3 and hilar neurons, suggesting that the small cluster of HSVGluR6-transduced cells induced an N-methyl-D-aspartate-dependent transsynaptic loss of non-transduced neurons.
患有严重颞叶癫痫的患者会在海马体的CA3区和门区失去神经元。通过用表达谷氨酸受体的GluR6红藻氨酸盐亚型的复制缺陷型单纯疱疹病毒（HSV）载体转导器官型海马切片培养物，也可诱导CA3区和门区神经元的丧失。在转导的成纤维细胞中，HSVGluR6表达一种分子量为115,000的蛋白质，该蛋白质与抗GluR6血清发生反应。将成纤维细胞暴露于HSVGluR6后，以前对红藻氨酸盐有抗性的细胞中出现了红藻氨酸盐依赖性毒性。将纳升量的重组HSV毒株微量注射到器官型海马切片培养物中，导致在微量注射部位出现局部转导和基因转移。尽管转导的神经元数量很少，但将100个HSVGluR6病毒粒子微量注射到CA3锥体层中会导致大量CA3锥体细胞和门区神经元丧失。当将100个HSVGluR6病毒粒子微量注射到CA1锥体层时，未观察到这种效应。河豚毒素或N-甲基-D-天冬氨酸受体拮抗剂可抑制CA3区和门区神经元的大量丧失，这表明少量HSVGluR6转导细胞诱导了非转导神经元的N-甲基-D-天冬氨酸依赖性跨突触丧失。
