Roy S J, Gilliham M, Berger B, Essah P A, Cheffings C, Miller A J, Davenport R J, Liu L-H, Skynner M J, Davies J M, Richardson P, Leigh R A, Tester M
Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge, CB2 3EA, UK.
Plant Cell Environ. 2008 Jun;31(6):861-71. doi: 10.1111/j.1365-3040.2008.01801.x. Epub 2008 Feb 13.
There is increasing evidence of the important roles of glutamate receptors (GLRs) in plant development and in adaptation to stresses. However, the studies of these putative ion channels, both in planta and in Xenopus oocytes, may have been limited by our lack of knowledge of possible GLR heteromer formation in plants. We have developed a modification of the single-cell sampling technique to investigate GLR co-expression, and thus potential heteromer formation, in single cells of Arabidopsis thaliana leaves. Micro-EXpression amplification (MEX) has allowed us to amplify gene transcripts from a single cell, enabling expression of up to 100 gene transcripts to be assayed. We measured, on average, the transcripts of five to six different AtGLRs in a single cell. However, no consistent patterns of co-expression or cell-type-specific expression were detected, except that cells sampled from the same plant showed similar expression profiles. The only discernible feature was the detection of AtGLR3.7 in every cell examined, an observation supported by GUS staining patterns in plants stably expressing promoter::uidA fusions. In addition, we found AtGLR3.7 expression in oocytes induces a Ba2+-, Ca2+- and Na+-permeable plasma membrane conductance.
越来越多的证据表明,谷氨酸受体(GLRs)在植物发育和适应胁迫中发挥着重要作用。然而,无论是在植物体内还是在非洲爪蟾卵母细胞中对这些假定离子通道的研究,可能都受到我们对植物中可能存在的GLR异源二聚体形成缺乏了解的限制。我们开发了一种单细胞采样技术的改进方法,以研究拟南芥叶片单细胞中的GLR共表达情况,进而研究潜在的异源二聚体形成。微表达扩增(MEX)使我们能够从单个细胞中扩增基因转录本,从而能够检测多达100种基因转录本的表达情况。我们平均在单个细胞中测量了五到六种不同AtGLR的转录本。然而,除了从同一植株中采样的细胞表现出相似的表达谱外,未检测到共表达或细胞类型特异性表达的一致模式。唯一可辨别的特征是在每个检测的细胞中都检测到了AtGLR3.7,在稳定表达启动子::uidA融合基因的植物中的GUS染色模式也支持了这一观察结果。此外,我们发现卵母细胞中AtGLR3.7的表达会诱导一种对Ba2+、Ca2+和Na+通透的质膜电导。