CNS MELATONIN TARGETS AND PHOTOPERIOD-INDUCED OBESITY

这项拨款提案的目的是开发新的范例，获得 新技术，并将其应用于当前研究的扩展 主题或与这些主题相关的新研究工作， 进一步发展PI的职业生涯。提出了三个具体目标， 每一个都有一系列的实验。 具体目标I：脂肪组织的神经支配在 季节性肥胖的光周期控制？实验被提议为 扩展了我们最近关于白色脂肪组织直接神经支配的报道 (WAT)交感神经系统（SNS）和我们发现的感觉 WAT的神经支配（见初步研究），以及每一个在 西伯利亚仓鼠季节性肥胖的光周期控制。四 问题将得到回答：1）短日照会导致身体减少吗？ 脂肪的发生独立于WAT？的SNS神经支配，2)是否 WAT的相对分离的SNS神经支配在CNS中会聚，3)什么 是WAT的感觉神经支配的程度，（4）什么是 WAT的感觉神经支配在短日照诱导的 减少身体脂肪？ 具体目标二：身体总量的关系是什么 脂肪和食物囤积的水平，以及感觉神经支配 在这段关系中的作用 西伯利亚仓鼠似乎 通过增加外部储存的能量来应对身体脂肪的减少， 食物储藏的形式。自然发生和实验诱导 体内脂肪的减少将被用来进一步测试相反的情况。 肥胖儿童体内脂肪总量与食物囤积水平的关系 西伯利亚仓鼠，以及WAT的感觉神经支配在这一过程中的作用。 关系食物囤积将使用模拟洞穴进行研究 系统三个问题将得到回答：1）做短日诱导 体内脂肪减少会刺激食物囤积？2)部分手术 脂肪切除术（脂肪去除）刺激食物囤积？，（3）角色是什么 短日照引起的WAT感觉神经支配的增加， 囤积食物 具体目标III：c-Fos活性是否可用于鉴定功能性 褪黑激素信号的接收/传输位点和其传出神经元 投影场 提出实验来检验这一假设， 刺激假定的接收/传输部位， 褪黑激素信号激活即刻早期基因c-Fos。这将是 通过使用皮下或皮下组织的定时侵入范例来完成， 生理相关褪黑激素脑内程序化递送 信号传递给切除松果体的西伯利亚仓鼠。 三个问题将是 回答：1）褪黑激素的全身输注是否激活了大脑中的神经元？ 西伯利亚仓鼠的三个大脑部位显示褪黑激素结合， 这些细胞的神经递质表型是什么2)在哪里 褪黑激素激活的细胞的传出投射野 并位于大脑中显示褪黑激素结合的三个部位，和3） 这些假定的传出投射野能否用逆行 纤维束示踪结合c-Fos免疫细胞化学技术？

The purpose of this grant proposal is to develop novel paradigms, acquire new techniques and apply them to either extensions of current research themes or to new research endeavors related to these themes in order to develop further the career of the PI. Three Specific Aims are proposed, each with a set of experiments. Specific Aim I: What is the role of the innervation of adipose tissue in the photoperiodic control of seasonal obesity? Experiments are proposed to extend our recent report of the direct innervation of white adipose tissue (WAT) by the sympathetic nervous system (SNS) and our finding of sensory innervation of WAT (see Preliminary Studies), and the role of each in the photoperiodic control of seasonal obesity in Siberian hamsters. Four questions will be answered: 1) Can the short day-induced decreases in body fat occur independently of the SNS innervation of WAT?, 2) Does the relatively separate SNS innervation of WAT converge in the CNS?, 3) What is the extent of the sensory innervation of WAT?, and 4) What is the functional role of the sensory innervation of WAT in the short day-induced decreases in body fat? Specific Aim II: What is the relationship between the amount of total body fat and level of food hoarding and what role does the sensory innervation of adlpose tissue play in this relationship? Siberian hamsters appear to respond to decreases in body fat by increasing externally stored energy in the form of a food hoard. Naturally occurring and experimentally induced decreases in body fat will be used to test further the inverse relationship between total body fat and the level of food hoarding in Siberian hamsters, and the role of the sensory innervation of WAT in this relationship. Food hoarding will be studied using a simulated burrow system. Three questions will be answered: 1) Do the short day-induced decreases in body fat stimulate food hoarding?, 2) Does partial surgical lipectomy (fat removal) stimulate food hoarding?, and 3)What is the role of the sensory innervation of WAT on the short day-induced increases in food hoarding? Specific Aim III: Can c-Fos activity be used to identify the functional sites of reception/transmission of melatonin signals and thefr efferent projection fields? Experiments are proposed to test the hypothesis that stimulation of the putative sites for the reception/transmission of melatonin signals activates the immediate-early gene c-Fos. This will be accomplished by using the timed infision paradigm for the subcutaneous or intracerebral programmed delivery of physiologically relevant melatonin signals to pinealectomized Siberian hamsters. Three questions will be answered: 1)Do systemic infusions of melatonin activate neurons in the three brain sites that show melatonin binding in Siberian hamsters and what are the neurotransmitter phenotypes of these cells?, 2) Where are the efferent projection fields of the cells that are activated by melatonin and located in the three brain sites that show melatonin binding?, and 3) Can these putative efferent projection fields be verified using retrograde tract tracing combined with c-Fos immunocytochemical techniques?