摘要
目的研究帕金森氏综合症的发病机理,评估wvGirk2对神经细胞生长发育和生命力的影响。方法给一类酪氨酸脱氢酶阳性的神经细胞(一种中枢神经源细胞),又称CAD细胞,引入由DNA编码的野生型和突变型离子通道。采用DNA克隆、免疫组化、WesternBlots免疫印迹等技术进行检测分析。用不同浓度(0.25μg/ml,0.5μg/ml,1.0μg/ml)的Girk2cDNA(对照组)和wvGirk2cDNA(观察组)质粒转染CAD细胞后,观察两组基因对转染CAD细胞的数目、蛋白质合成、神经突生长等指标的影响作用;并且观察MK801和Kir2.3对wvGirk2的抑制作用。结果高浓度wvGirk2转染的CAD细胞存活数目减少到约60%的单纯Girk2转染细胞数目;低浓度wvGirk2没有引起细胞死亡但是减少转染基因的蛋白质产物,且神经突生长也受wvGirk2影响;MK801和Kir2.3对wvGirk2有抑制作用。结论wvGirk2基因在weaver动物中有一种特殊功能,阻断wvGirk2通道能够防止细胞死亡。离子通道的存在和wvGirk2的转染可能对wvGirk2细胞的命运有显著影响。
Objective To study the etiology of Parkinson′ s Disease and to assess the impact of wvGirk2 on neuronal development and vitality. Methods To introduce DNA encoding the wild- type and mutant channel into immortalized tyrosine hydroxylase- positive CNS- derived neurons, CAD cells. DNA clone, Immunostaining and Western Blots were used. Three different concentrations of Girk2 and wvGirk2 expression plasmids were transfected: 0.25g/ml, 0.5g/ml, or 1.0g/ml. Observed the number of transfected cycling cells, protein synthesis and neurites growth between two groups. Results Transfection of cycling CAD cells with high concentration of wvGirk2 reduced the number of cells to about 60% of Girk2- transfected cells. Low concentration of wvGirk2 did not cause cell death but reduced the protein products of transfected genes. Neurite growth is also affected by wvGirk2. MK- 801 and Kir2.3 can alter the effect of wvGirk2. Conclusion The study results support the finding that wvGirk2 has a gain- of- function effect in weaver animals in that blocking the wvGirk2 channel results in the rescue of cells from death. Our data also suggest that the presence of channels and the level of wvGirk2 may have a significant impact on the fate of cells containing wvGirk2. [
出处
《中国航天医药杂志》
2003年第4期7-12,共6页
Medical Journal of CASE
