大鼠嘌呤P2Y2受体在神经系统分布的实验研究

目的探讨P2Y2受体在脊髓、背根神经节和坐骨神经的表达和具体分布，研究三磷酸腺苷（adenosine triphosphate．ATP）对神经再生的作用机制提供理论基础。方法将6只SD乳鼠的脊髓、背根神经节和坐骨神经标本置于含有0．1％二乙基焦碳酸的4%多聚甲醛液中快速固定、石蜡包埋、超薄切片。实验组：根据P2Y2受体mRNA的碱基序列，制作非放射性标记的核苷酸探针，原位杂交后将切片置于含有NBT溶液（50mg／ml硝基兰四唑溶于二甲基甲酰胺）、BCIP溶液（75mg／ml溴绿一吲哚磷酸溶于二甲基酰胺）的检测缓冲液中染色，镜下观察P2Y2受体的具体分布。对照组：切片刚不含探针的杂交缓冲液覆盖、染色观察。结果实验组：脊髓灰质前角神经元胞浆中可见P2Y2受体的表达，呈弱阳性；背根神经节雪旺细胞的胞浆和胞核中均可见P2Y2受体的表达，且呈强阳性；对照组：无P2Y2受体表达。实验组与对照组坐骨神经原位杂交尢P2Y2受体的表达。结论P2Y2受体主要分布在脊髓、背根神经节，细胞外ATP可以通过P2Y2受体作用于脊髓和背根神经节细胞。

EXPERIMENTAL STUDY ON EXPRESSION OF THE PURINE P2Y2 RECEPTOR IN RAT NERVE SYSTEM

Objective To study the distribution of P_2Y_2 receptor in spine cord, dorsal root ganglia and sciatic nerve in rat, and to provide the basis for clarifying the mechanism of the effect of adenosine triphosphate(ATP) on the peripheral nerve regeneration.Methods Six specimens of the spine cord, dorsal root ganglia and sciatic nerve from SD rats were fixed rapidly in 4% paraformaldehyde which included DEPC, imbedded by paraffin and made into ultrathin section. According to the sequence of P_2Y_2 receptor's gene, DNA needle was adopted to detect the distribution of P_2Y_2 receptor by hybridization technique in section under the light microscope after they had been stained in NBT liquid(50 mgml) and BCIP liquid (75 mgml). In the control group, the ultrathin section was only covered with hybridism buffer solution. The result of staining was observed. Results Hybridization in section showed that P_2Y_2 receptor was distributed mainly in the anterior horn cell of spine cord gray matter and Schwann cell of the dorsal root ganglia. No P_2Y_2 receptor was observed in the sciatic nerve of both groups. Conclusion P_2Y_2 receptor is located mainly in the spine cord and the dorsal root ganglia. Extracellular ATP can affect the cell of spine cord, dorsal root ganglia through P_2Y_2 receptor.