神经生长因子mRNA在大鼠脊髓和脊神经节的表达——原位杂交研究

The Study of Expressions of Nerve Growth Factor mRNA in the Rat Spinal Cord and Spinal Ganglion with in Situ Hybridization Histochemistry

我们以 SD大鼠坐骨神经为材料 ,在 NGF- c DNA文库建立的基础上 ,人工合成神经生长因子引物 ,并用 PCR地高辛标记法标记 NGF探针 ,采用原位分子杂交组织化学方法 ( ISHH) ,观察 NGF- m RNA神经生长基因表达细胞在大鼠腰段脊髓和脊神经节内的分布。结果发现在大鼠坐骨神经损伤模型腰段脊髓横切面的前角、侧角及腰背根神经节均有 NGF基因的表达细胞 ,蓝色反应物弥散性分布于胞浆内 ,呈细小颗粒状或长柱状。损伤侧要强于未损伤侧 ,并对其杂交信号进行定量分析 ,结果显示在大鼠坐骨神经损伤模型术后第 5天、第 10天及第 15天 ,脊髓前角运动神经元 ,侧角交感神经元、背根节感觉神经元内的杂交信号增强 ,表明损伤的早、中期 NGF- m RNA表达量增加。

On the basis of NGF cDNA library of rat sciatic nerve tissues in regeneration, then the probe was labeled with digoxin by means of PCR. NGF clones. The distribution of NGF mRNA gene expressing cells in the rat lumbar spinal cord and spinal ganglion was studied by in situ hybridization histochemistry method. The results showed that NGF gene expressing cells were in anterior horn, lateral horn of spinal cord and lumbar spinal ganglion in rat sciatic nerve injury model. Blue reactants spread all over the cytoplasm and presented fine granular or longer pillar with stronger in injured sides of spinal cord than in non injured sides, and the signal of hybridization was analysed by morphometry. The results showed that the signal of hybridization was strong in the motor neuron of anterior horn, sympathetic neuron of lateral horn, sensory neuron of the spinal ganglion on the 5th, 10th and 15th days after rat sciatic nerve injury had been performed. It was demonstrated that the expression of NGF mRNA increased at the early, middle stage of nerve injury. This article discussed the factors of physics and chemistry in nerve regeneration.