海马结构内PENK和PDYN基因表达之间关系的探讨

The restrictive relationship between PENK and PDYN gene expression in hippocampus of rat

关键词:  寡核苷酸  PENK基因  PDYN基因  微渗透泵  海马

Key words:  oligodeoxynucleotides  PENK gene  PDYN gene  miniosmotic pump  hippocampus

摘要:

讨海马结构内PENK和PDYN基因表达之间的关系。方法：合成一段与PENK mRNA起始部位互补的39 mer Antisense，同时分别合成PENK mRNA的Sense序列和Missense序列。将三段寡核苷酸硫代化后，用微渗透泵通过脑内给药的双通不锈钢管，分别将其恒速持续注入清醒自由活动大鼠的一侧腹侧海马结构内，并用原位杂交技术观察海马内PENK mRNA和PDYN mRNA的表达。结果：灌注Antisense侧海马内PENK mRNA的表达较对侧明显降低，而PDYN mRNA却较对侧明显增高；灌注Sense 和Missense 的大鼠两侧海马内PENK mRNA和PDYN mRNA的表达未见明显差别，提示Antisense选择性阻断 PENK mRNA 表达后，同侧PENK mRNA表达降低的同时PDYN mRNA表达明显增加；同时发现对侧PENK mRNA表达增加的同时PDYN mRNA的表达明显降低，并且反义寡核苷酸的作用具有序列特异性。结论：PENK mRNA的表达能抑制 PDYN mRNA的表达，二者具有相互制约关系。

Abstract:

To study the restrictive relationship between PENK and PDYN gene expression in hippocampus of rat 39 mer antisense being complementary to PENK mRNA translation initiation sequence was synthetized. Meanwhile the sense and missense sequence of PENK mRNA were synthetized respectively. After all oligodeoxynucleotides (ODNs) were phosphorothioated,they were continuously infused into the unilateral ventral hilus in rats in doses of 0.5μl/h, 20nmol/day for 5 days by miniosmotic pump and cannula.The expression of PENK and PDYN mRNA in hippocampus were observed by in situ hybridization method.The result showed that the expression of PENK mRNA in the infused antisense side of the hippocampus was significantly lower than that in the other side and the expression of PDYN mRNA in the infused side is higher than that in the other side.The expression of PENK mRNA and PDYN mRNA exhibited no significant difference in both sides of the hippocampus of the rats infused with sense and missense. It was suggested that after antisense selectively blocked PENK mRNA expression, PENK mRNA expression at the same side decreased while PDYN mRNA expression increased.But at the opposite side,PENK mRNA expression increased while PDYN mRNA expression decreased. The action of antisense is specific in sequence. Our work demonstrated that the expression of PENK mRNA could inhibit the expression of PDYN mRNA and both of them could condition each other.