Visualization of Sensory Neurons and Their Projections in an Upper Motor Neuron Reporter Line
Visualization of Sensory Neurons and Their Projections in an Upper Motor Neuron Reporter Line
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DOI：
10.1371/journal.pone.0132815
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发表时间：
2015-07-29
期刊：
影响因子：
3.7
通讯作者：
Oezdinler, P. Hande
中科院分区：
文献类型：
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作者：
Genc, Baris;Lagrimas, Amiko Krisa Bunag;Oezdinler, P. Hande
外周神经系统轴突和细胞体的可视化对于理解它们的发育、目标识别和整合到复杂的电路中是重要的。许多研究使用了蛋白质基因产物（PGP） 9.5[又名PGP]。泛素羧基末端水解酶L1 （UCHL1）的表达作为标记感觉神经元及其轴突的标志物。在UCHL1启动子控制下，绿色荧光蛋白（eGFP）的表达增强在UCHL1-eGFP报告系中稳定且持久。除了运动皮质脊髓运动神经元和脊髓抗退行性脊髓运动神经元的遗传标记外，我们还报道了周围神经系统的神经元在UCHL1-eGFP报告系中也被荧光标记。eGFP的表达在胚胎时期开启，并持续到成年期，从而可以详细研究体内所有感觉神经元的细胞体、轴突和目标神经支配模式。此外，可视化同一动物的感觉神经元和运动神经元提供了许多优势。在本报告中，我们使用了UCHL1-eGFP报告细胞系在两种不同的疾病范式：糖尿病和运动神经元疾病。在高脂饮食诱导的糖尿病模型中，感觉轴突中eGFP的表达有助于确定表皮神经纤维密度的变化。我们的发现证实了以前的研究，并表明需要超过5个月的时间来进行明显的皮肤去神经支配。UCHL1-eGFP与hSOD1（G93A）小鼠杂交产生肌萎缩性侧索硬化症hSOD1(G93A)-UeGFP报告系，发现感觉神经系统缺陷，尤其是疾病终末期。我们的研究不仅强调了ALS疾病的复杂性，也揭示了UCHL1-eGFP报告线将是一个有价值的工具，可以在许多疾病的背景下观察和研究感觉神经系统发育和退化的各个方面。
Visualization of peripheral nervous system axons and cell bodies is important to understand their development, target recognition, and integration into complex circuitries. Numerous studies have used protein gene product (PGP) 9.5 [a.k.a. ubiquitin carboxy-terminal hydrolase L1 (UCHL1)] expression as a marker to label sensory neurons and their axons. Enhanced green fluorescent protein (eGFP) expression, under the control of UCHL1 promoter, is stable and long lasting in the UCHL1-eGFP reporter line. In addition to the genetic labeling of corticospinal motor neurons in the motor cortex and degeneration-resistant spinal motor neurons in the spinal cord, here we report that neurons of the peripheral nervous system are also fluorescently labeled in the UCHL1-eGFP reporter line. eGFP expression is turned on at embryonic ages and lasts through adulthood, allowing detailed studies of cell bodies, axons and target innervation patterns of all sensory neurons in vivo. In addition, visualization of both the sensory and the motor neurons in the same animal offers many advantages. In this report, we used UCHL1-eGFP reporter line in two different disease paradigms: diabetes and motor neuron disease. eGFP expression in sensory axons helped determine changes in epidermal nerve fiber density in a high-fat diet induced diabetes model. Our findings corroborate previous studies, and suggest that more than five months is required for significant skin denervation. Crossing UCHL1-eGFP with hSOD1(G93A) mice generated hSOD1(G93A)-UeGFP reporter line of amyotrophic lateral sclerosis, and revealed sensory nervous system defects, especially towards disease end-stage. Our studies not only emphasize the complexity of the disease in ALS, but also reveal that UCHL1-eGFP reporter line would be a valuable tool to visualize and study various aspects of sensory nervous system development and degeneration in the context of numerous diseases.