Detloff Megan Ryan, Fisher Lesley C, Deibert Rochelle J, Basso D Michele
Center for Brain and Spinal Cord Repair, School of Allied Medical Professions, The Ohio State University, USA.
J Vis Exp. 2012 Apr 4(62):e3247. doi: 10.3791/3247.
Spinal cord injury (SCI) impairs sensory systems causing allodynia. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia(3). Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.
脊髓损伤(SCI)会损害感觉系统,导致痛觉过敏。为了确定痛觉过敏的细胞和分子原因,需要在大鼠SCI模型中进行灵敏且有效的感觉测试。然而,直到最近,还没有一种单一的测试方法在SCI中得到验证,因此各实验室尚未采用标准化方法。此外,现有的测试方法无法在急性情况下或存在严重运动障碍时实施,从而阻碍了对SCI诱导的痛觉过敏发展过程的研究(3)。在此,我们介绍两种使用von Frey Hair(VFH)单丝的经过验证的感觉测试方法,这些方法可量化SCI后触觉感觉阈值的变化。一种测试是成熟的上下测试,在长期测试时,该测试在不同的SCI严重程度下均显示出高灵敏度和特异性。另一种测试是新开发的背部VFH测试,当痛觉过敏出现且运动恢复之前,该测试可在SCI后急性应用。每个VFH单丝接触后爪皮肤时会施加校准力,直到其弯曲。在上下方法中,在足底L5皮节上使用交替的较高或较低力的VFH来确定屈肌退缩阈值。依次施加更高的力,直到出现退缩,然后使用较低力的VFH,直到退缩停止。触觉阈值反映了在50%的刺激中引发退缩所需的力。对于新测试,在检查者支撑大鼠时,将每个VFH应用于爪的背部L5皮节。VFH刺激按力的升序进行,直到在给定力下3次应用中至少2次导致爪退缩。触觉感觉阈值是66%的时间内引发退缩的最低力。描述了适应、测试和评分程序。定义了需要重新测试的异常试验和典型试验。动物使用得到了俄亥俄州立大学动物护理和使用委员会的批准。