Department of Orthopedics, The Third Affiliated Hospital, Suzhou University, Changzhou, China.
Acta Histochem. 2011 Jan;113(1):43-8. doi: 10.1016/j.acthis.2009.08.003. Epub 2009 Sep 23.
We have used semi-quantitative RT-PCR, Western blot, and immunofluorescence imaging approaches to detect the expression levels of Slit2 and its receptor Robo1 in the rat spinal cord after traumatic lesions. Our results revealed that both the mRNA and protein levels of Slit2 were up-regulated in the injured spinal cord. The Slit2 expression level was increased at day 7 until day 14, and then returned to normal level at day 21 after injury. A double-immunolabelling study showed that Slit2 and neurofilament (NF) proteins were both localized in neurons of spinal corda cinerea. Slit2 immunopositivity was detected in neuronal plasma membranes but not in the axonal fibers. In contrast, the immunolabelling of Robo1 in the normal spinal cord was at a low level, mostly in the neurons of spinal corda cinerea, and remained unchanged at all time points following spinal cord injury (SCI). The regulation levels of Slit2 and Robo1 after traumatic lesions in the rat spinal cord are different. Our results indicate that Slit2-Robo1 might not be involved in the inhibitory environment after SCI.
我们使用半定量 RT-PCR、Western blot 和免疫荧光成像方法来检测创伤性损伤后大鼠脊髓中 Slit2 及其受体 Robo1 的表达水平。我们的结果表明，Slit2 的 mRNA 和蛋白水平在损伤的脊髓中均上调。Slit2 的表达水平在第 7 天增加到第 14 天，然后在损伤后第 21 天恢复正常水平。双重免疫标记研究表明，Slit2 和神经丝（NF）蛋白均定位于脊髓灰质中的神经元中。Slit2 免疫阳性反应可检测到神经元质膜，但不存在于轴突纤维中。相比之下，Robo1 在正常脊髓中的免疫标记水平较低，主要位于脊髓灰质中的神经元中，并且在脊髓损伤（SCI）后所有时间点均保持不变。大鼠脊髓创伤性损伤后 Slit2 和 Robo1 的调节水平不同。我们的结果表明，Slit2-Robo1 可能不参与 SCI 后的抑制环境。
