Adenovirus-mediated expression of a reporter gene in thalamocortical cocultures.
Adenovirus-mediated expression of a reporter gene in thalamocortical cocultures.
复制标题
丘脑皮质共培养物中腺病毒介导的报告基因表达。
DOI：
10.1016/0006-8993(95)01101-3
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发表时间：
1995
期刊：
影响因子：
2.9
通讯作者：
Angelides,KJ
中科院分区：
文献类型：
--
作者：
Wilkemeyer,MF;Angelides,KJ
丘脑和皮层外植体的器官型共培养最近被用来研究哺乳动物新皮层中丘脑皮层轴突网络的发育。为了探索遗传操作器官型外植体的可能性，用重组腺病毒Adv/RSVβgal感染大鼠丘脑皮质（TC）共培养物。皮质外植体的感染导致报告基因（β-半乳糖苷酶）的长期表达（2周），外植体的结构完整性没有显著改变。通过将腺病毒显微注射到皮层外植体中，获得了对报告基因表达的显著程度的空间控制。还分析了从丘脑外植体到感染（n = 116）和对照皮质（n = 120）外植体的Dil标记的轴突投射。TC长入感染或对照皮质外植体的范围或程度无显著差异。用Adv/RSVβgal病毒也能有效感染丘脑外植体。虽然感染的丘脑外植体的TC向内生长的模式和程度与对照相似，但存活的感染的丘脑外植体的百分比降低。这些实验是将来使用重组腺病毒和器官型共培养物进行研究的必要前提。这些共培养物的遗传操作应该能够使用适合于直接操作和观察的系统来解剖参与哺乳动物新皮层中轴突网络发展的蛋白质。
Organotypic cocultures of thalamic and cortical explants have recently been used to study the development of the thalamocortical axonal network in the mammalian neocortex. To explore the possibility of genetically manipulating organotypic explants, rat thalamocortical (TC) cocultures were infected with the recombinant adenovirus, Adv/RSVβgal. Infection of the cortical explants resulted in long-term expression (2 weeks) of the reporter gene (β-galactosidase) with no significant alterations to the structural integrity of the explants. By micro-injecting the adenoviruses into cortical explants a significant degree of spatial control over reporter gene expression was obtained. Dil-labeled axonal projections from thalamic explants into infected (n = 116) and control cortical (n = 120) explants were also analyzed. There was no significant difference in the extent or degree of TC ingrowth into infected or control cortical explants. Thalamic explants were also efficiently infected with the Adv/RSVβgal virus. While the pattern and extent of TC ingrowth from infected thalamic explants was similar to controls, the percentage of viable, infected thalamic explants was decreased. These experiments were necessary precursors for future studies using recombinant adenoviruses and organotypic cocultures. Genetic manipulation of these cocultures should enable the dissection of proteins involved in the development of axonal networks in the mammalian neocortex, using a system amenable to direct manipulation and observation.