Getchell T V, Grillo M, Tate S S, Urade R, Teeter J, Margolis F L
Department of Neuroscience, Roche Research Center, Nutley, N.J. 07110.
Neurochem Res. 1990 Apr;15(4):449-56. doi: 10.1007/BF00969932.
We demonstrate that poly (A+)RNA isolated from catfish barbels directs the expression of functional amino acid taste receptors in the Xenopus oocyte. The activity of these receptors is monitored in ovo by the two electrode voltage clamp technique. Specific conductance changes recorded in response to amino acid stimulation are analogous to those recorded electrophysiologically from intact catfish barbels. These responses exhibit specificity, reproducibility, rapid onset and termination, and desensitization to repetitive stimulation. A functional assay system that encompasses the full complement of transduction events from the ligand-receptor interaction to subsequent conductance changes is necessary to identify molecular components responsible for these events. Our results demonstrate that the Xenopus oocyte can be used to characterize and identify clones coding for amino acid taste receptors analogous to its use in studying receptor molecules for other neuroactive compounds.
我们证明,从鲶鱼触须分离的聚腺苷酸(poly(A+))RNA可指导非洲爪蟾卵母细胞中功能性氨基酸味觉受体的表达。这些受体的活性通过双电极电压钳技术在卵内进行监测。响应氨基酸刺激记录的特定电导变化类似于从完整鲶鱼触须电生理记录的变化。这些反应具有特异性、可重复性、快速起始和终止以及对重复刺激的脱敏作用。一个涵盖从配体-受体相互作用到后续电导变化的完整转导事件的功能测定系统对于鉴定负责这些事件的分子成分是必要的。我们的结果表明,非洲爪蟾卵母细胞可用于表征和鉴定编码氨基酸味觉受体的克隆,这类似于其在研究其他神经活性化合物的受体分子中的应用。