Identification of thermosensory and olfactory neuron-specific genes via expression profiling of single neuron types
Identification of thermosensory and olfactory neuron-specific genes via expression profiling of single neuron types
复制标题
DOI：
10.1016/j.cub.2004.12.030
复制
发表时间：
2004-12-29
期刊：
影响因子：
9.2
通讯作者：
Sengupta, P
中科院分区：
文献类型：
--
作者：
Colosimo, ME;Brown, A;Sengupta, P
大部分C。秀丽线虫感觉神经元类型由单个双侧神经元对组成，并且对一组独特的感觉刺激作出反应。虽然已经在正向遗传筛选中鉴定了个体感觉神经元类型的发育和功能所需的基因，但这些方法不太可能鉴定出突变时导致微妙或多效性表型的基因。在这里，我们描述了一种互补的方法来确定感觉神经元类型特异性基因，通过微阵列分析使用RNA从分类AWB嗅觉和AFD热敏神经元。这些基因的子集的表达模式进一步在体内验证。通过该分析鉴定的基因编码7-跨膜受体、激酶和核因子，包括编码高度保守的腊肠犬蛋白的同源物[1]的EST-I。TTX-1在包括AFD神经元在内的感觉神经元亚群中表达，并受TTX-1 OTX同源结构域蛋白调节[2]。在热梯度上，ESTA-1突变体不能抑制嗜冷驱动，但在培养温度下继续跟踪等温线，这代表了这些AFD介导的行为的第一次遗传分离。单个神经元类型的表达谱分析提供了一种快速、强大和无偏见的方法，用于识别神经元特异性基因，然后可以在体内研究其功能。
Most C. elegans sensory neuron types consist of a single bilateral pair of neurons, and respond to a unique set of sensory stimuli. Although genes required for the development and function of individual sensory neuron types have been identified in forward genetic screens, these approaches are unlikely to identify genes that when mutated result in subtle or pleiotropic phenotypes. Here, we describe a complementary approach to identify sensory neuron type-specific genes via microarray analysis using RNA from sorted AWB olfactory and AFD thermosensory neurons. The expression patterns of subsets of these genes were further verified in vivo. Genes identified by this analysis encode 7-transmembrane receptors, kinases, and nuclearfactors including dac-1, which encodes a homolog of the highly conserved Dachshund protein [1]. dac-1 is expressed in a subset of sensory neurons including the AFD neurons and is regulated by the TTX-1 OTX homeodomain protein [2]. On thermal gradients, dac-1 mutants fail to suppress a cryophilic drive but continue to track isotherms at the cultivation temperature, representing the first genetic separation of these AFD-mediated behaviors. Expression profiling of single neuron types provides a rapid, powerful, and unbiased method for identifying neuron-specific genes whose functions can then be investigated in vivo.