PCR标记探针原位检测培养的海马神经细胞BDNF mRNA的表达

利用现成的BDNF成熟基因片段和上下游引物，用PCR直接扩增得到DIG标记的BDNF探针。用此探针原位检测海马神经细胞BDNFmRNA表达状况．通过设置阴性对照组和竞争实验组，说明杂文反应是特异的．原位杂交法观察神经细胞经药物处理后BDNFmRNA的变化，与文献报道用Northern法的结果一致。另外观察到BDNFmRNA表达差异，不仅存在于海马不同区，在同一区的细胞群也不同。这种原位杂交方法，操作相对简便，但灵敏度还不能检测BDNPmRNA在胶质细胞中的表达。

IN SITU DETECTION OF BDNF mRNA IN CULTURED HIPPOCAMPAL NEURONS WITH PROBE LABELED BY PCR

The DIG labeled BDNF probe was synthesized by PCR technique from mature hurnan BDNF gene constructed in plasmid M13mp 18 with specific primers. Witll this probe we detected the expression of BDNF mRNA in situ in cultured neurons. The specificity of the hybridization was dernonstrated by negative experirnents and competitive experiments. The changes of BDNF mRNA amou nt in neurons after treatment with drugs were consistent with those of Northern blot analyses reported by others. And we observed that besides its regional distribution in hippocampus the expressions of BDNF mRNA were not the same in neurons in the CA1 or DG region. This in situ hybridization rnethod is relatively simple, but its sensi-tivity needs to be improved since it is unable to detect the basal expression of BDNF mRNA in situ in astrocytes by this methed.