Chen J, Kelz M B, Zeng G, Sakai N, Steffen C, Shockett P E, Picciotto M R, Duman R S, Nestler E J
Laboratory of Molecular Psychiatry and Center for Genes and Behavior, Departments of Psychiatry and Neurobiology, Yale University School of Medicine, New Haven, Connecticut 06508, USA.
Mol Pharmacol. 1998 Sep;54(3):495-503. doi: 10.1124/mol.54.3.495.
Several inducible gene expression systems have been developed in vitro in recent years to overcome limitations with traditional transgenic mice. One of these, the tetracycline-regulated system, has been used successfully in vivo. Nevertheless, concerns remain about the ability of this system to direct high levels of transgene expression in vivo and to enable such expression to be turned on and off effectively. We report here the generation, using a modified tetracycline-regulated system under the control of the neuron-specific enolase promoter, of several lines of mice that direct transgene expression to specific brain regions, including the striatum, cerebellum, CA1 region of the hippocampus, or deep layers of cerebral neocortex. Transgene expression in these mice can be turned off completely with low doses of doxycycline (a tetracycline derivative) and driven to very high levels in the absence of doxycycline. We demonstrate this tissue-specific, inducible expression for three transgenes: those that encode luciferase (a reporter protein) or DeltaFosB or the cAMP-response element binding protein (CREB) (two transcription factors). The various lines of transgenic mice demonstrate an inducible system that generates high levels of transgene expression in specific brain regions and represent novel and powerful tools with which to study the functioning of these (or potentially any other) genes in the brain.
近年来，为克服传统转基因小鼠的局限性，人们在体外开发了几种诱导型基因表达系统。其中之一，四环素调控系统，已在体内成功应用。然而，对于该系统在体内指导高水平转基因表达以及有效开启和关闭这种表达的能力，仍存在担忧。我们在此报告，利用神经元特异性烯醇化酶启动子控制下的改良四环素调控系统，培育出了几条转基因小鼠品系，这些品系可将转基因表达导向特定脑区，包括纹状体、小脑、海马体CA1区或大脑新皮质深层。在这些小鼠中，低剂量强力霉素（一种四环素衍生物）可完全关闭转基因表达，而在没有强力霉素的情况下，转基因表达可被驱动至非常高的水平。我们对三种转基因证明了这种组织特异性的诱导表达：编码荧光素酶（一种报告蛋白）或DeltaFosB或环磷酸腺苷反应元件结合蛋白（CREB）（两种转录因子）的转基因。各种转基因小鼠品系展示了一种诱导系统，该系统可在特定脑区产生高水平的转基因表达，代表了研究这些（或潜在任何其他）基因在大脑中功能的新颖而强大的工具。
