The mouse tissue plasminogen activator gene 5' flanking region directs appropriate expression in development and a seizure-enhanced response in the CNS.
The mouse tissue plasminogen activator gene 5' flanking region directs appropriate expression in development and a seizure-enhanced response in the CNS.
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发表时间：
1994-11
期刊：
影响因子：
4.6
通讯作者：
Pamela M. Carroll;Stella E. Tsirka;William G. Richards;M. A. Frohman;Sidney Strickland
中科院分区：
文献类型：
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作者：
Pamela M. Carroll;Stella E. Tsirka;William G. Richards;M. A. Frohman;Sidney Strickland
组织纤溶酶原激活剂 (t-PA) 是一种分泌型丝氨酸蛋白酶，与发育的多个方面有关。在成年大鼠大脑中，t-PA 的转录是诱导神经元可塑性治疗后海马体的立即早期反应。为了研究控制该基因转录的序列元件，使用原位分析来定义妊娠中期胚胎中 t-PA 的时间和空间表达模式。然后产生携带与大肠杆菌lacZ基因连接的t-PA 5'侧翼序列的转基因小鼠。含有 4 kb 侧翼序列 (4.0TAMGAL) 的构建体主要向通过原位分析显示高水平 t-PA mRNA 的相同组织赋予 β-半乳糖苷酶活性。在胚胎第 8.5 天 (E8.5) 至 13.5 天 (E13.5) 的 4.0TAMGAL 胚胎中，观察到的大部分表达位于神经外胚层衍生的组织中。 β-半乳糖苷酶活性首先在中脑和间脑的限制性神经节中检测到，分别位于 E8.5 和 E9.5。在 E10.5，在神经嵴衍生的脑神经和背根神经节中观察到转基因表达，但在基板衍生的脑神经中没有观察到转基因表达。从 E10.5 到 E13.5，在中脑、脊髓、神经视网膜和发育中的嗅觉系统的有丝分裂后神经元中观察到 β-半乳糖苷酶活性。在正在进行组织重塑的区域（例如耳廓、胡须毛囊和四肢）也检测到了 β-半乳糖苷酶活性。在成年小鼠中，lacZ 在海马体中表达，并且发现这种表达在 CA3 巨锥体神经元癫痫发作时增强。这些结果强化了t-PA在神经发生和形态发生中发挥作用的概念，并确定了在发育和中枢神经系统中指导其转录调节的启动子区域。
Tissue plasminogen activator (t-PA) is a secreted serine protease implicated in multiple aspects of development. In the adult rat brain, transcription of t-PA is an immediate-early response in the hippocampus following treatments that induce neuronal plasticity. To study the sequence elements that govern transcription of this gene, in situ analysis was used to define t-PA's temporal and spatial expression pattern in midgestation embryos. Transgenic mice were then generated carrying t-PA 5' flanking sequences linked to the E. coli lacZ gene. Constructs containing 4 kb of the flanking sequences (4.0TAMGAL) confer beta-galactosidase activity mostly to the same tissues that exhibit high levels of t-PA mRNA by in situ analysis. In 4.0TAMGAL embryos from embryonic day 8.5 (E8.5) to 13.5 (E13.5), the majority of expression observed is localized to neural ectoderm-derived tissues. beta-galactosidase activity is first detected in restricted neuromeres in the midbrain and diencephalon, at E8.5 and E9.5 respectively. At E10.5, transgene expression is observed in neural crest-derived cranial nerves and dorsal root ganglia, but not placode-derived cranial nerves. From E10.5 to E13.5, beta-galactosidase activity is observed in postmitotic neurons of the midbrain, spinal cord, neural retina and the developing olfactory system. beta-galactosidase activity is also detected in areas undergoing tissue remodeling such as the pinna of the ear, whisker follicles and the limbs. In adult mice, lacZ is expressed in the hippocampus and this expression was found to be enhanced upon seizure in the giant pyramidal neurons of CA3. These results reinforce the concept that t-PA plays a role in neurogenesis and morphogenesis, and identifies the promoter region that directs its transcriptional regulation both in development and in the CNS.