5-羟色胺转运体基因异体表达模型的建立

ESTABLISHMENT OF HETEROLOGOUS EXPRESSION MODEL OF hSERT IN XENOPUS LAEVIS OOCYTES

目的:探讨建立5-羟色胺转运体(5-HTT)基因异体表达模型的可行性,且为进一步探讨5-羟色胺重摄取的动力过程和条件以及其功能的调控机制奠定基础。方法:利用体外转录cRNA技术将克隆至pOTV中的5-HTT的cDNA转录、合成5HTT的cRNA,通过显微注射技术将该cRNA注入成熟雌性非洲爪蟾卵母细胞的胞质中,使其表达以建立5-羟色胺转运体(SERT)的异体表达模型,并用电压钳技术检测其转运功能。结果:爪蟾卵母细胞可被用做5-HTT的异体表达模型,其转运功能呈浓度依赖性并具饱和现象,转运过程可被特异阻断剂De-sipramine阻断。结论:非洲爪蟾卵母细胞可作为5-羟色胺等单胺类神经递质转运体的异体表达系统,为进一步研究转运体蛋白的功能和调控提供了有效工具。

Aim： To determine the feasibility of establishing the heterologous expression model of human- serotonine transporter（hSERT or 5-HTT）. Mothods： cRNA of SERT was transcribed from cDNA, which was cloned in the pOTV vector. Each oocyte of mature xenopus laevis was injected with transcribed cRNA in vim and incubated at room temperature for 4-9 days. Recording the current induced by 5-HT with voltage clamp technique tested the function of the expressed 5-HT transporter. Results： The transporter current could be observed in Ringer＇s .solution containing 5-HT, and the 5-HT induced current were concentration-dependent. Norepinephrine and dopamine could not induce the transporter current while the 5-HT induced current could be specifically inhibited by 5- HTT blocker, desipramine. Conclusion： The results demonstrate that the heterologous expression product in xenopus lavevis oocytes is human 5-HT transporter.