GENES CONTROLLING SOCIAL BEHAVIOR IN ANTS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 A.具体目标 原提案的具体目标： 1：确定预测任务行为的蜜蜂基因的火蚁同源物。 2：将已知的社会任务特定脑区大小的变化与基因表达的差异联系起来。 3：沉默特定基因的表达，以测试社交行为的丧失。 修改具体目标1，以回应2004年8月10日总结声明中的批评意见： 1.确定预测任务行为的蜜蜂基因的火蚁同源物，并确定社交中使用的化学信号。 B.研究和成果 1.涉及任务行为和社会交流的基因。我们试图通过原位杂交来定位蛋白激酶G表达的脑区，但得到了不一致的结果。我们在洛桑火蚁文库中鉴定了11个化学感觉蛋白(CSP)序列。CSP1对应于我们先前发现的一个序列，该序列是工蜂触角中主要的低分子蛋白质。我们克隆并在大肠杆菌中表达了重组CSP1，并对其进行了层析纯化。通过结合时NPN荧光增强检测到重组CSP1与N-苯基萘胺(NPN)结合。我们测得NPN与蝗虫CSP-SG4的亲和力约为10？M，而NPN与CSP-SG4的亲和力为4？M。我们将提取的角质层碳氢化合物加入CSP1-NPN复合体中，发现提取物的一个或多个成分取代了NPN在CSP1上的结合部位。因此，角质层碳氢化合物提取物中含有一个或多个信号分子。 C.意义 昆虫模型系统关系到人类的健康，昆虫模型系统具有一些优点，包括避免在脊椎动物身上进行实验和简化基因操作。 D.计划 我们将用免疫细胞化学的方法确定PKG在火蚁脑中的表达区域。角质层碳氢化合物提取物的CSP1结合成分将被鉴定。将进行PKG的RNAi研究。 E.出版物：无 F.项目生成的资源：无

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A. Specific Aims Specific aims in original proposal: 1: Identify fire ant homologs of bee genes predictive of task behavior. 2: Correlate known social task-specific size changes in brain regions with differences in gene expression. 3: Silence expression of specific genes to test for loss of social behavior. Modification of specific aim 1 in response to critiques in Summary Statement of 8-10-2004: 1. Identify fire ant homologs of bee genes predictive of task behavior, and identify chemical signals used in social communication. B. Studies and Results 1. Genes involved in task behavior and social communication. We attempted to locate the brain region of protein kinase G expression by in situ hybridization, but inconsistent results were obtained. We identified 11 chemosensory protein (CSP) sequences in the Lausanne Fire Ant Library. CSP1 corresponded to a sequence which we previously found to be the major low molecular weight protein in worker antennae. We cloned and expressed recombinant CSP1 in E. coli and purified it by chromatography. The recombinant CSP1 binds to N-phenylnaphthylamine (NPN), as detected by enhancement of NPN fluorescence upon binding. We measured the affinity as about 10 ¿M, which compares with 4 ¿M for NPN binding to locust CSP-sg4. We added extracted cuticular hydrocarbons to the CSP1-NPN complex and found that one or more components of the extract displaced NPN from its binding site on CSP1. Thus, one or more signalling molecules are contained in the cuticular hydrocarbon extract. C. Significance Insect model systems are relevant to human health, and insect model systems have some advantages, including avoidance of experimentation on vertebrates and the simplicity of gene manipulations. D. Plans We will identify the region of expression of PKG in fire ant brains using immunocytochemistry. The CSP1-binding components of the cuticular hydrocarbon extract will be identified. RNAi studies of PKG will be done. E. Publications: none F. Project-generated resources: none