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目的 :建立研究神经生理及药理的卵母细胞表达模型 ,并对鸡脑mRNA在卵母细胞上表达的氨基酸类受体进行研究。方法 :采用盐酸胍或异硫氰酸胍法抽提总RNA ,经寡聚脱氧胸苷 (oligo -dT)纤维素柱纯化得mRNA。mRNA微注射到卵母细胞内进行表达 ,以双电极电压钳技术研究表达的受体。结果 :模型能明显表达外源性mRNA表达的受体 ,而阴性对照无表达。在卵母细胞膜上表达的γ -氨基丁酸 (γ -aminobutyricacid ,GABA) ,甘氨酸(Glycine ,Gly)及红藻氨酸 (Kainate ,KA)受体 ,施加GABA ,Gly及KA均能引起一剂量依赖的内向电流 ,相应的拮抗剂可阻断其电流。剂量效应曲线显示GABA及KA诱导电流的半有效浓度 (EC5 0 )分别为 2 .9× 10 -5mol/L及 6 .3× 10 -5mol/L。结论 :卵母细胞可明显表达鸡脑mRNA编码的KA ,Gly及GABA受体 ,其药理特性与天然受体相似 ,是一个良好的神经生理及药理研究模型。
OBJECTIVE: To establish a neuropsychological and pharmacological oocyte expression model and to study the amino acid receptors expressed on the oocytes of chicken brain mRNA. Methods: Total RNA was extracted with guanidine hydrochloride or guanidium isothiocyanate and purified by oligo-dT cellulose column. The mRNA was microinjected into oocytes for expression and the expressed receptors were studied by bipolar voltage clamp technique. Results: The model can express the receptor of exogenous mRNA expression, but not the negative control. GABA, GABA, KA and KA expressed on oocyte membrane could induce a dose of GABA, Gly and KA Dependent on the inward current, the corresponding antagonist can block its current. The dose-response curve showed that the EC50 of GABA and KA-induced currents were 2.9 × 10 -5 mol / L and 6.3 × 10 -5 mol / L, respectively. CONCLUSION: Oocytes can express KA, Gly and GABA receptors encoded by chicken brain mRNA and their pharmacological properties are similar to those of natural receptors. This is a good neurophysiological and pharmacological model.