Widespread neuron-specific transgene expression in brain and spinal cord following synapsin promoter-driven AAV9 neonatal intracerebroventricular injection
Widespread neuron-specific transgene expression in brain and spinal cord following synapsin promoter-driven AAV9 neonatal intracerebroventricular injection
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DOI：
10.1016/j.neulet.2014.05.044
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发表时间：
2014-07-25
影响因子：
2.5
通讯作者：
Isacson, Ole
中科院分区：
文献类型：
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作者：
McLean, Jesse R.;Smith, Gaynor A.;Isacson, Ole
腺相关病毒（AAV）基因转移具有治疗广泛的神经退行性疾病的巨大希望。AAV 9血清型穿过血脑屏障，并且与其他血清型相比显示出增强的转导效率，从而在需要整体转基因表达时提供有利的靶向。新生儿静脉或脑室内（i. c. v.）重组AAV 9（rAAV 9）的递送最近已被证明对于建模和治疗神经变性疾病的几种啮齿动物模型是有效的，然而，该技术与可变的细胞向性相关，使得定制的基因转移成为挑战。在目前的研究中，我们采用人突触蛋白1（hSYN 1）基因启动子驱动绿色荧光蛋白（GFP）的神经元特异性表达后，新生儿i. c. v.注射rAAV 9小鼠。我们观察到广泛的GFP表达在整个大脑，脊髓，周围神经和神经节在6周龄的神经元。GFP表达的区域特异性定量显示黑质网状部的高神经元转导率（43.9 ± 5.4%），运动皮质（43.5 +/- 3.3%），海马（43.1 ± 2.7%）、小脑（29.6 ± 2.3%）、颈脊髓（24.9 ± 3.9%）和腹内侧纹状体（16.9 ± 4.3%）。我们发现14.6 +/- 2.2%的支配腓肠肌的神经肌肉接头显示GFP免疫反应性。在几种神经元亚型中发现了GFP表达，包括黑质酪氨酸羟化酶（TH）阳性多巴胺能细胞、纹状体多巴胺和cAMP调节的神经元磷蛋白（DARPP-32）阳性神经元和胆碱乙酰转移酶（ChAT）阳性运动神经元。这些结果建立在当代基因转移技术的基础上，表明hSYN 1启动子可以与rAAV 9一起用于在整个神经系统中驱动强大的神经元特异性转基因表达。(C)2014作者出版社：Elsevier爱尔兰Ltd.
Adeno-associated viral (AAV) gene transfer holds great promise for treating a wide-range of neurodegenerative disorders. The AAV9 serotype crosses the blood brain barrier and shows enhanced transduction efficiency compared to other serotypes, thus offering advantageous targeting when global transgene expression is required. Neonatal intravenous or intracerebroventricular (i.c.v.) delivery of recombinant AAV9 (rAAV9) have recently proven effective for modeling and treating several rodent models of neurodegenerative disease, however, the technique is associated with variable cellular tropism, making tailored gene transfer a challenge. In the current study, we employ the human synapsin 1 (hSYN1) gene promoter to drive neuron-specific expression of green fluorescent protein (GFP) after neonatal i.c.v. injection of rAAV9 in mice. We observed widespread GFP expression in neurons throughout the brain, spinal cord, and peripheral nerves and ganglia at 6 weeks-of-age. Region-specific quantification of GFP expression showed high neuronal transduction rates in substantia nigra pars reticulata (43.9 +/- 5.4%), motor cortex (43.5 +/- 3.3%), hippocampus (43.1 +/- 2.7%), cerebellum (29.6 +/- 2.3%), cervical spinal cord (24.9 +/- 3.9%), and ventromedial striatum (16.9 4.3%), among others. We found that 14.6 +/- 2.2% of neuromuscular junctions innervating the gastrocnemius muscle displayed GFP immunoreactivity. GFP expression was identified in several neuronal sub-types, including nigral tyrosine hydroxylase (TH)-positive dopaminergic cells, striatal dopamine-and cAMP-regulated neuronal phosphoprotein (DARPP-32)-positive neurons, and choline acetyltransferase (ChAT)-positive motor neurons. These results build on contemporary gene transfer techniques, demonstrating that the hSYN1 promoter can be used with rAAV9 to drive robust neuron-specific transgene expression throughout the nervous system. (C) 2014 The Authors. Published by Elsevier Ireland Ltd.