Liu Xiaorong, Feng Liang, Shinde Ishan, Cole James D, Troy John B, Saggere Laxman
Department of Ophthalmology, Northwestern University, Chicago, IL, USA.
Department of Mechanical and Industrial Engineering, University of Illinois at Chicago, Chicago, IL, USA.
Neurol Res. 2020 May;42(5):379-386. doi: 10.1080/01616412.2020.1733322. Epub 2020 Feb 26.
: Rodent models of optic nerve crush (ONC) have often been used to study degeneration and regeneration of retinal ganglion cells (RGCs) and their axons as well as the underlying molecular mechanisms. However, ONC results from different laboratories exhibit a range of RGC injury with varying degree of axonal damage. We developed instrumented tweezers to measure optic nerve (ON) crush forces in real time and studied the correlation between RGC axon loss and force-impulse, the product of force and duration, applied through the instrumented tweezers in mice.: A pair of standard self-closing #N7 tweezers were instrumented with miniature foil strain gauges at optimal locations on both tweezers' arms. The instrumented tweezers were capable of recording the tip closure forces in the form of voltages, which were calibrated through load cells to corresponding tip closure forces over the operating range. Using the instrumented tweezers, the ONs of multiple mice were crushed with varied forces and durations and the axons in the immunostained sections of the crushed ONs were counted.: We found that the surviving axon density correlated with crush force, with longer duration and stronger crush forces producing consistently more axon damage.: The instrumented tweezers enable a simple technique for measurement of ONC forces in real-time for the first time. Using the instrumented tweezers, experimenters can quantify crush forces during ONC to produce consistent and predictable post-crush cell death. This should permit future studies a way to produce nerve damage more consistently than is available now.
视神经挤压(ONC)啮齿动物模型常被用于研究视网膜神经节细胞(RGCs)及其轴突的退化与再生以及潜在的分子机制。然而,不同实验室的ONC结果显示出一系列不同程度轴突损伤的RGC损伤情况。我们开发了带仪器的镊子以实时测量视神经(ON)挤压力,并研究了RGC轴突损失与通过带仪器的镊子施加于小鼠的力-冲量(力与持续时间的乘积)之间的相关性。
一对标准的自动闭合#N7镊子在其双臂的最佳位置安装了微型箔式应变片。带仪器的镊子能够以电压形式记录尖端闭合力,通过称重传感器在工作范围内将其校准为相应的尖端闭合力。使用带仪器的镊子,以不同的力和持续时间挤压多只小鼠的视神经,并对挤压后的视神经免疫染色切片中的轴突进行计数。
我们发现,存活的轴突密度与挤压力相关,持续时间越长、挤压力越强,产生的轴突损伤就越严重。
带仪器的镊子首次实现了一种简单的实时测量ONC力的技术。使用带仪器的镊子,实验者可以在ONC过程中量化挤压力,以产生一致且可预测的挤压后细胞死亡情况。这应该为未来的研究提供一种比目前更能一致地造成神经损伤的方法。