Ogurusu T, Taira H, Shingai R
Department of Computer and Information Science, Faculty of Engineering, Iwate University, Japan.
J Neurochem. 1995 Sep;65(3):964-8. doi: 10.1046/j.1471-4159.1995.65030964.x.
We identified GABAA receptor subunits in rat retina using PCR. The high degree of conservation among previously described members of ligand-gated anion channels in transmembrane domains was used to design degenerate sense and antisense oligonucleotides. These oligonucleotides were used as primers for PCR, which was applied to the rat retina cDNA. Analysis of clones derived from the PCR amplification identified the GABAA alpha 1, beta 1, beta 3, and gamma 2 subunits and the glycine alpha 1 subunit. In addition, two clones closely related to the human GABAA rho-subunit class were obtained. Molecular cloning revealed one of them as the rat counterpart of the human rho 2 subunit. Northern blot analysis demonstrated the expression of mRNAs for rho subunits in retina. These results further support the hypothesis that bicuculline-insensitive GABA channels in rat retina are comprised of rho subunits.
我们使用聚合酶链反应（PCR）在大鼠视网膜中鉴定了GABAA受体亚基。利用跨膜结构域中先前描述的配体门控阴离子通道成员之间的高度保守性来设计简并正义和反义寡核苷酸。这些寡核苷酸用作PCR引物，应用于大鼠视网膜cDNA。对PCR扩增得到的克隆进行分析，鉴定出了GABAAα1、β1、β3和γ2亚基以及甘氨酸α1亚基。此外，还获得了两个与人类GABAAρ亚基类别密切相关的克隆。分子克隆显示其中一个是人类ρ2亚基的大鼠对应物。Northern印迹分析表明视网膜中存在ρ亚基的mRNA表达。这些结果进一步支持了大鼠视网膜中荷包牡丹碱不敏感的GABA通道由ρ亚基组成的假说。
