Kang Xiaoning, Wen Jingkun, Wang Xianghai, Pan Mengjie, Zhang Weiwei, Zhan Xiaoduo, Liu Zhongying, Wu Wutian, Guo Jiasong
Department of Histology and Embryology, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2013 Apr;33(4):463-8.
To quantitatively analyze the temporal and spatial pattern of RhoA expression in injured spinal cord of adult mice.
A spinal cord transection model was established in adult mice. At 1, 3, 7, 14, 28, 56 and 112 days after the surgery, the spinal cords were dissected and cryosectioned for RhoA/NF200, RhoA/GFAP, RhoA/CNPase or RhoA/IBA1 double fluorescent immunohistochemistry to visualize RhoA expressions in the neurons, astrocytes, oligodendrocytes and microglia. The percentages as well as the immunostaining intensities of RhoA-positive cells in the parenchymal cells were quantitatively analyzed.
RhoA was weakly expressed in a few neurons and oligodendrocytes in normal spinal cord. After spinal cord injury, the percentage of RhoA-positive cells and RhoA expression intensity in the spinal cord increased and peaked at 7 days post injury (dpi) in neurons, oligodendrocytes and astrocytes, followed by a gradual decrease till reaching a low level at 112 dpi. In the microglia, both the RhoA-positive cells and RhoA expression intensity reached the maximum at 14 dpi and maintained a high level till 112 dpi.
Traumatic spinal cord injury can upregulate RhoA expression in the neurons as well as all the glial cells in the spinal cord. RhoA expression patterns vary with post-injury time, location and among different parenchymal cells in the injured spinal cord.
定量分析成年小鼠脊髓损伤后RhoA表达的时空模式。
在成年小鼠中建立脊髓横断模型。术后1、3、7、14、28、56和112天,解剖脊髓并进行冷冻切片,用于RhoA/NF200、RhoA/GFAP、RhoA/CNPase或RhoA/IBA1双重荧光免疫组织化学,以观察神经元、星形胶质细胞、少突胶质细胞和小胶质细胞中RhoA的表达。对实质细胞中RhoA阳性细胞的百分比和免疫染色强度进行定量分析。
正常脊髓中,少数神经元和少突胶质细胞中RhoA呈弱表达。脊髓损伤后,脊髓中RhoA阳性细胞的百分比和RhoA表达强度增加,并在损伤后7天(dpi)在神经元、少突胶质细胞和星形胶质细胞中达到峰值,随后逐渐下降,直到112 dpi时降至低水平。在小胶质细胞中,RhoA阳性细胞和RhoA表达强度均在14 dpi时达到最大值,并维持在高水平直至112 dpi。
创伤性脊髓损伤可上调脊髓中神经元以及所有胶质细胞中RhoA的表达。RhoA的表达模式随损伤后时间、位置以及损伤脊髓中不同实质细胞而变化。