SYSTEMATIC AND CELLULAR INDICIES OF STRESS IN OBESITY

描述(改编自应用程序)：长期目标是更好地 了解脊椎动物对压力的动态平衡反应。它 被提议使用人类病理生理学的大鼠模型，特别是 肥胖，以调查全身和细胞的反应性 在压力下发生了变化。睡眠不足将被用作压力 关于遗传性肥胖(fa/fa)和瘦(Fa/？)的范例扎克老鼠。 在这个模型中，PI将检查神经内分泌和心血管 涉及四个动态平衡系统的压力指数：(1) 下丘脑-垂体-甲状腺(HPT)轴；(3)交感肾上腺(SA)轴。 系统，包括内分泌胰腺；和(4)血流动力学参数 心血管(CV)系统。具体目标是：(1) 通过确定神经内分泌反应性的特征和比较 组织和血浆HPA轴激素(CRF、ACTH、催乳素和 皮质酮)；HPT轴(TRF、TSH和甲状腺激素，包括 总成分和游离成分)和SA系统激素(NE、E、DA；和 儿茶酚胺合成酶)；(2)测定血流动力学 心血管系统通过测量平均动脉压的响应性， 心率、呼吸频率和心电波形； 通过确定细胞对压力的变化来测量细胞对压力的反应 应激相关基因(热激基因)的表达；及(4) 测定关键代谢限速酶活性的变化 作为细胞碳流变化指标的关键代谢途径 和能源利用。目标1和目标3的实验方法将 使用免疫分析，Western印迹，Northern印迹， 免疫细胞化学、原位杂交和高效液相色谱检测特异性 指定分析的本地化和定量测定。 目标2将通过植入无线电遥测设备来实现 测量心血管参数。 关于应激如何影响患有精神障碍的大鼠模型，我们知之甚少。 肥胖是许多人类的特征。有效的假设是 以下是：考虑到在施加强有力的应激源时观察到的发病率 对于正常的瘦肉动物来说，长期的假设是， 有其自身的潜在病理也会导致发病率， 结果显著缩短了体内平衡的时间 反应性与瘦小的动物相比，反应性将 在规模上比瘦小的动物更大。这些预测 观察结果将与健康状况的整体下降相一致 压力过大的动物，表明在体内平衡方面，肥胖者 表型明显处于更大的生理苦恼中。

Description (Adapted from Application): The long-term goal is to better understand the homeostatic responsiveness of vertebrates to stress. It is proposed to use a rat model of human pathophysiology, specifically obesity, to investigate how systemic and cellular responsiveness is altered during stress. Sleep deprivation will be employed as the stress paradigm on the genetically obese (fa/fa) and lean (Fa/?) Zucker rat. With this model, the PI will examine neuroendocrine and cardiovascular indices of stress involving four homeostatic systems: (1) the hypothalamic-pituitary-thyroid (HPT) axis; (3) the sympathoadrenal (SA) system, including endocrine pancreas; and (4) hemodynamic parameters of the cardiovascular (CV) system. The specific aims are: (1) to characterize and compare neuroendocrine responsiveness by determining tissue and plasma levels of HPA axis hormones (CRF, ACTH, prolactin, and corticosterone); HPT axis (TRF, TSH, and thyroid hormones, including both total and free components); and SA system hormones (NE, E, DA; and catecholamine synthesizing enzymes); (2) to determine the hemodynamic responsiveness of the CV system by measuring mean arterial pressure, heart rate, respiratory rate, and electrocardiogram waveforms; (3) to measure cellular responsiveness to stress by determining changes in expression of stress-related genes (heat-shock genes); and (4) to measure changes in activities of rate-limiting enzymes of key metabolic of key metabolic pathways as an index of change in cellular carbon flow and energy utilization. Experimental approaches for aims 1 and 3 will be to use immunoassay, Western blots, Northern blots, immunocytochemistry, in situ hybridization, and HPLC for specific localization and quantitative determination of the designated analyses. Aim 2 will be accomplished by implantation of radiotelemetry devices to measure cardiovascular parameters. Little is known about how stress affects a rat model with a disorder characteristic of many humans, obesity. The working hypotheses is as follows: Given the morbidity observed when a potent stressor is applied chronically to normal lean animals, it is hypothesized that obesity, with its own underlying pathologies that also lead to morbidity, will result in a significantly shorter time course of homeostatic responsiveness compared to lean animals and that the responsiveness will be greater in magnitude compared to lean animals. These predicted observations will be consistent with the overall decline of the health of stressed animals, indicating that homeostatically, the obese phenotype is in markedly greater physiological distress.