Identification of a Drosophila glucose receptor using Ca2+ imaging of single chemosensory neurons.
Identification of a Drosophila glucose receptor using Ca2+ imaging of single chemosensory neurons.
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DOI：
10.1371/journal.pone.0056304
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Amrein H
中科院分区：
文献类型：
--
作者：
Miyamoto T;Chen Y;Slone J;Amrein H
化学感受细胞对食物化合物的评价对于动物做出适当的摄食决定是必不可少的。在果蝇Drosophila melanogaster中，由味觉受体（Gr）蛋白大家族成员组成的多聚体受体检测结构多样的化学物质。推定的糖和苦味受体在味觉感受器的味觉受体神经元（GRN）的不同子集中表达，从而分别将不同的味觉品质赋予糖和苦味化合物。在这里，我们报告了一种钙离子成像方法，允许关联的配体介导的反应，一个单一的GRN。我们发现，不同的甜神经元表现出不同的响应曲线时，刺激各种糖，同样，不同的苦神经元表现出不同的响应曲线时，刺激一组苦的化学物质。这些观察结果表明，在一个味觉通道内的单个神经元分别由不同的甜味和苦味受体库表示。此外，我们采用这种新的方法来确定葡萄糖作为糖受体Gr61a的主要配体，它不仅在经典化学感受器的甜味感受神经元中表达，而且在两个超敏感的非典型味觉感受器神经元中表达。因此，单细胞Ca 2+成像可以作为一个强大的工具，以确定孤儿Gr蛋白的配体。
Evaluation of food compounds by chemosensory cells is essential for animals to make appropriate feeding decisions. In the fruit fly Drosophila melanogaster, structurally diverse chemicals are detected by multimeric receptors composed of members of a large family of Gustatory receptor (Gr) proteins. Putative sugar and bitter receptors are expressed in distinct subsets of Gustatory Receptor Neurons (GRN) of taste sensilla, thereby assigning distinct taste qualities to sugars and bitter tasting compounds, respectively. Here we report a Ca2+ imaging method that allows association of ligand-mediated responses to a single GRN. We find that different sweet neurons exhibit distinct response profiles when stimulated with various sugars, and likewise, different bitter neurons exhibit distinct response profiles when stimulated with a set of bitter chemicals. These observations suggest that individual neurons within a taste modality are represented by distinct repertoires of sweet and bitter taste receptors, respectively. Furthermore, we employed this novel method to identify glucose as the primary ligand for the sugar receptor Gr61a, which is not only expressed in sweet sensing neurons of classical chemosensory sensilla, but also in two supersensitive neurons of atypical taste sensilla. Thus, single cell Ca2+ imaging can be employed as a powerful tool to identify ligands for orphan Gr proteins.
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