Pheromone regulation of gene expression in the brain

描述(申请人提供)：我们将把一个新的模型(蜜蜂)与强大的新基因组资源(cDNA微阵列和即将完成的全基因组序列)相结合，以研究信息素如何调节大脑中的基因表达。蜜蜂内分泌调节行为发育的方式与哺乳动物生殖发育的调节方式非常相似，受引子信息素的调节，而蜜蜂蜂后下颌信息素(QMP)是为数不多的影响内分泌调节生理和行为发育的引子信息素之一。我们将：1)确定Kr-H1的表达与信息素调控行为的关系，Kr-ht是在高级大脑中心发现的第一个信息素调控基因，它是一种转录因子，并通过两种社会相关方式进行调控。我们将通过一系列社会、行为和药物操作来检验这一假设，即QMP抑制Kr-H1表达与这种信息素对内分泌介导的行为成熟的抑制作用有关。2)用免疫细胞化学和果蝇强大的MARCM技术以及蜜蜂(与Tzumin Lee)互补的方法，确定Kr-H1表达对脑细胞神经解剖和突触结构的影响。3)确定Kr-ht转录调控的下游靶点。作为一种转录因子，Kr-H1可能通过控制其他基因的表达来发挥作用，这些基因可能在神经元重塑或调节觅食行为中发挥更直接的功能作用。我们将使用多管齐下的方法来鉴定这些基因，包括：鉴定蜜蜂和果蝇Kr-hl蛋白的共同结合序列；生物信息学(与Hugh Robertson)；通过肽序列和染色质重塑分析表征与Kr-hl相关的蛋白质；蜜蜂和果蝇的微阵列实验；以及染色质免疫沉淀(与Craig Mizzen)这项研究的主要意义是它将提高我们对化学沟通如何影响神经和行为可塑性的分子理解。

DESCRIPTION (provided by applicant): We will couple a novel model (honey bee) with powerful new genomic resources (cDNA microarray and a soon-to-be-completed complete genome sequence) to study how pheromones regulate gene expression in the brain. Endocrine-mediated behavioral development in the bee is regulated in a manner strikingly similar to the regulation of mammalian reproductive development, by primer pheromones, and honeybee queen mandibular pheromone (QMP) is one of the few chemically characterized primer pheromones known to affect endocrine-mediated physiological and behavioral development. We will: 1) Determine how Kr-hl expression is related to pheromone regulation of behavior, Kr-ht, the first pheromone regulated gene identified in a higher brain center, is a transcription factor and is regulated in two socially relevant ways. We will test the hypothesis that QMP repression of Kr-hl expression is related to this pheromone's inhibitory effects on endocrine-mediated behavioral maturation with a set of social, behavioral, and pharmacological manipulations. 2) Determine the effect of Kr-hl expression on brain cell neuroanatomy and synaptic structure using immunocytochemistry and the powerful MARCM technique in Drosophila and complementary approaches in bee (with Tzumin Lee). 3) Identify downstream targets of Kr-ht transcriptional regulation. As a transcription factor, Kr-hl presumably functions by controlling expression of other genes that would play more direct functional roles in neuronal remodeling or regulating foraging behavior. We will use a multi-pronged approach to identify these genes involving, identification the consensus binding sequence of the bee and Drosophila Kr-hl proteins; bioinformatics (with Hugh Robertson); characterization of proteins associated with Kr-hl by peptide sequencing and chromatin remodeling assays; microarray experiments in both bees and flies; and chromatin immunoprecipitation (with Craig Mizzen) The principal significance of this research is that it will improve our molecular understanding of how chemical communication influences neural and behavioral plasticity.