论文部分内容阅读
为了观察β-1,4-半乳糖基转移酶-I(β-1,4-galactosyltransferase-I,β-1,4-GalT-I)在正常和钳夹伤后大鼠坐骨神经中表达的变化,本研究采用RT-PCR方法,从小鼠坐骨神经中特异性地扩增β-1,4-GalT-I cDNA片段并将其克隆到pGEM-T载体。采用体外转录的方法合成地高辛标记的正、反义β-1,4-GalT-I RNA探针。通过原位杂交和图像分析的方法,观察β-1,4-GalT-I mRNA在正常和夹伤大鼠坐骨神经中的表达及其变化。结果表明:β-1,4-GalT-I mRNA在大鼠坐骨神经的髓鞘中表达,在夹伤坐骨神经后1~2d,β-1,4-GalT-I mRNA在坐骨神经的表达最高,于夹伤后1周开始下降,在夹伤后1个月恢复至正常水平。上述结果提示坐骨神经夹伤后β-1,4-GalT-I mRNA的表达发生变化,并且主要表达在坐骨神经的Schwann细胞。本研究结果为进一步分析β-1,4-GalT-I在周围神经再生中的调控机制奠定了基础。
In order to observe the changes of the expression of β-1,4-galactosyltransferase-I (β-1,4-GalT-I) in rat sciatic nerve after normal and clamp injury In this study, β-1,4-GalT-I cDNA fragment was amplified from mouse sciatic nerve by RT-PCR and cloned into pGEM-T vector. Digoxigenin labeled positive and antisense β-1,4-GalT-I RNA probes were synthesized by in vitro transcription. The expression and changes of β-1,4-GalT-I mRNA in the sciatic nerve of normal and injured rats were observed by in situ hybridization and image analysis. The results showed that β-1,4-GalT-I mRNA was expressed in the myelin sheath of rat sciatic nerve. The expression of β-1,4-GalT-I mRNA in sciatic nerve was the highest at 1 ~ 2 d after sciatic nerve injury 1 week after injury began to decline, 1 month after the injury returned to normal levels. The above results suggest that the expression of β-1,4-GalT-I mRNA changes after sciatic nerve injury and is mainly expressed in Schwann cells of the sciatic nerve. The results of this study provide a basis for further analysis of the regulation of β-1,4-GalT-I in peripheral nerve regeneration.