SOCIAL AND NEUROENDOCRINE REGULATION OF THE PERIOD GENE

调节昼夜节律的分子也被认为在不同的时间尺度上影响过程的功能。该项目的目标是测试该模型的一个重要预测：鉴于行为和分子数据表明蜜蜂的行为成熟和行为昼夜节律之间存在关联，那么已知影响行为成熟率的社会和神经内分泌因素也应该影响per表达。将进行以下六项调查。1. 描述蜜蜂的每个基因：我们已经生成了一个全长cDNA序列，也对基因组克隆进行了部分测序，并将进行比较分析（与H. Robertson）。2. 确定蜜蜂大脑中per mRNA水平是否在行为成熟过程中发生变化。在蜂巢里工作的小蜜蜂没有行为上的昼夜节律，而在花上觅食的老蜜蜂有；大脑中per mRNA水平的变化与这种行为成熟有关。相关分析为Aims 4-6中的处理实验提供了基线数据。3. 定位并测量脑PER水平。初步免疫细胞化学结果表明，per mRNA的成熟变化反映在蛋白水平上。继续的分析将与原位杂交相结合，以提供额外的确认（与S. Fahrbach和K. Siwicki）。这项相关研究的结果也将用于为目标4-6提供基线。4. 确定影响行为成熟的社会环境的变化是否也会影响大脑中的每一种表达。我们将充分利用一套独特的强大的行为操作来分离实际年龄，行为成熟阶段和经验，所有这些都是与彼此高度相关的蜜蜂一起工作的。初步结果支持这一假设。5. 确定已知影响行为成熟的幼年激素和章鱼胺操作是否会导致节律性运动活动和6。每个表达式。运动活动的初步结果支持这一假设。我们假设幼激素类似物和章鱼胺处理会增加每一个表达，而allatectomy和章鱼胺拮抗剂处理会降低它。这项研究的主要意义在于，它首次尝试通过实验确定时钟基因的表达是否依赖于比那些与昼夜节律严格相关的更广泛的影响。

Molecules that regulate circadian rhythmicity are also thought to influence the functioning of processes at different temporal scales. The goal of this project is to test an important prediction of this model: given that behavioral and molecular data suggest an association between behavioral maturation and behavioral circadian rhythms in the honey bee, then social and neuroendocrine factors known to influence rate of behavioral maturation should also influence per expression. The following six lines of inquiry will be pursued. 1. Characterize the bee per gene: We have generated a full-length cDNA sequence and also have partially sequenced a genomic clone and will do comparative analyses (with H. Robertson). 2. Determine whether per mRNA levels in the bee brain change during behavioral maturation. Young bees, which work in the hive, have no behavioral circadian rhythms while older bees, which forage on flowers, do; per mRNA levels in the brain change in association with this behavioral maturation. This correlative analysis provides baseline data for the treatment experiments in Aims 4-6. 3. Localize and measure levels of brain PER. Preliminary immunocytochemical results indicate that maturational changes in per mRNA are reflected at the protein level. Continued analyses will be coupled with in situ hybridization to provide additional confirmation (with S. Fahrbach and K. Siwicki). Results of this correlative study will also be used to provide a baseline for Aims 4-6. 4. Determine whether changes in social environment, known to influence behavioral maturation, also influence per expression in the brain. We will take full advantage of a uniquely powerful set of behavioral manipulations to dissociate chronological age, stage of behavioral maturation, and experience, all while working with bees that are highly related to each other. Preliminary results support this hypothesis. 5. Determine whether juvenile hormone and octopamine manipulations, known to influence behavioral maturation, cause changes in both rhythmic locomotor activity and 6. per expression. Preliminary results for locomotor activity support this hypothesis. We hypothesize that juvenile hormone analog and octopamine treatments will increase per expression while allatectomy and octopamine antagonist treatment will decrease it. The principal significance of this research is that it attempts to experimentally determine for the first time whether the expression of a clock gene is dependent upon a broader array of influences than those strictly associated with circadian rhythmicity.