Interdepartmental Neuroscience Program, Yale University, New Haven, Connecticut 06520-8074, USA.
J Neurosci. 2010 Mar 31;30(13):4562-72. doi: 10.1523/JNEUROSCI.6357-09.2010.
A central question in insect chemoreception is whether signaling occurs via G-proteins. Two families of seven-transmembrane-domain chemoreceptors, the odor (Or) and gustatory receptor (Gr) families, have been identified in Drosophila (Clyne et al., 1999, 2000; Vosshall et al., 1999). Ors mediate odor responses, whereas two Grs, Gr21a and Gr63a, mediate CO2 response (Hallem et al., 2004; Jones et al., 2007; Kwon et al., 2007). Using single-sensillum recordings, we systematically investigate the role of Galpha proteins in vivo, initially with RNA interference constructs, competitive peptides, and constitutively active Galpha proteins. The results do not support a role for Galpha proteins in odor sensitivity. In parallel experiments, manipulations of Galpha(q), but not other Galpha proteins, affected CO2 response. Transient, conditional, and ectopic expression analyses consistently supported a role for Galpha(q) in the response of CO2-sensing neurons, but not odor-sensing neurons. Genetic mosaic analysis confirmed that odor responses are normal in the absence of Galpha(q). Ggamma30A is also required for normal CO2 response. The simplest interpretation of these results is that Galpha(q) and Ggamma30A play a role in the response of CO2-sensing neurons, but are not required for Or-mediated odor signaling.
昆虫化学感受中的一个核心问题是信号是否通过 G 蛋白传递。在果蝇中已经鉴定出了两类具有七个跨膜结构域的化学感受器,即气味(Or)和味觉受体(Gr)家族(Clyne 等人,1999 年,2000 年;Vosshall 等人,1999 年)。Ors 介导气味反应,而两个 Grs,Gr21a 和 Gr63a,介导 CO2 反应(Hallem 等人,2004 年;Jones 等人,2007 年;Kwon 等人,2007 年)。我们使用单感器记录,系统地研究了 Galpha 蛋白在体内的作用,最初使用 RNAi 构建体、竞争肽和组成性激活的 Galpha 蛋白。结果不支持 Galpha 蛋白在嗅觉敏感性中的作用。在平行实验中,Galpha(q)的操纵,但不是其他 Galpha 蛋白的操纵,影响了 CO2 反应。瞬时、条件和异位表达分析一致支持 Galpha(q)在 CO2 感受神经元反应中的作用,但不支持嗅觉感受神经元的作用。遗传镶嵌分析证实,在缺乏 Galpha(q)的情况下,嗅觉反应正常。Ggamma30A 也需要正常的 CO2 反应。对这些结果的最简单解释是,Galpha(q)和 Ggamma30A 在 CO2 感受神经元的反应中起作用,但不是 Or 介导的气味信号所必需的。