Pheromone regulation of gene expression in the brain

描述（申请人提供）：我们将结合一个新的模型（蜜蜂）和强大的新的基因组资源（cDNA微阵列和即将完成的全基因组序列）来研究信息素如何调节大脑中的基因表达。蜜蜂内分泌介导的行为发育与哺乳动物生殖发育的调节方式惊人地相似，由引物信息素调节，而蜂王下颌信息素（QMP）是已知的少数具有化学特征的影响内分泌介导的生理和行为发育的引物信息素之一。我们将：1)确定Kr-hl表达与信息素调节行为的关系，Kr-ht是第一个在高级脑中心发现的信息素调节基因，是一种转录因子，并通过两种社会相关方式进行调节。我们将通过一系列社会、行为和药理学操作来验证QMP抑制Kr-hl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的假设。2)利用免疫细胞化学和强大的MARCM技术在果蝇和蜜蜂中确定Kr-hl表达对脑细胞神经解剖学和突触结构的影响（与Tzumin Lee）。3)确定Kr-ht转录调控的下游靶点。作为一种转录因子，Kr-hl可能通过控制其他基因的表达来发挥作用，这些基因在神经元重塑或调节觅食行为中发挥更直接的功能作用。我们将采用多管齐下的方法来鉴定这些基因，鉴定蜜蜂和果蝇Kr-hl蛋白的一致结合序列；生物信息学（与休·罗伯逊合作）；通过肽测序和染色质重塑测定来鉴定与Kr-hl相关的蛋白质；蜜蜂和苍蝇的微阵列实验；这项研究的主要意义在于，它将提高我们对化学通讯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.