INTERACTION OF INTERLEUKIN 1 WITH CRF NEURONS

白细胞介素 1 与 CRF 神经元的相互作用

• 批准号: 3416200
• 负责人: JOHN A OLSCHOWKA
• 金额: $ 15.28万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3416200
• 关键词: blood brain barrier; central nervous system; cholera toxin; corticotropin releasing factor; cytokine receptors; electron microscopy; gene expression; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; interleukin 1; laboratory rat; messenger RNA; microscopy; neuroendocrine system; neurons; oncogenes; physiologic stressor; psychoneuroimmunology; stress

The body displays a remarkable ability to maintain a relatively constant internal milieu when faced with increasing physical or psychological demands. A variety of autonomic and endocrine responses, coordinated in the CNS, serve to adapt the body to stress. In addition, the immune system plays an equal part in facing a variety of antigens. Increasing evidence has suggested that these three regulatory systems of the body are in constant communication. It is well established that glucocorticoids are elevated during an immune response and in turn inhibit the immune response. This feedback loop is thought to begin among immune cells that release a multitude of cytokines, including interleukin-1 (IL-1). IL-1 is a low molecular weight polypeptide that is believed to have endocrinologic, neurologic and immunologic functions. Within the CNS IL-1 is known to cause the release of corticotropin releasing factor (CRF) into the portal vessels, induce fever, cause anorexia, increase slow-wave sleep, increase central autonomic activity, etc. While the preoptic nucleus has been implicated as a site of action of IL-1 in inducing fever and CRF release, in general we do not know of the sites of action of IL-1 within the CNS. We now propose to study several aspects of IL-1 interactions with cells of the central CRF neuroendocrine system. The specific aims we will attempt to complete are: 1) To determine the distribution of activated neurons within the CNS following stimulation with IL-1beta. We will use a new neuroanatomic technique for functionally mapping the neurons activated by IL-1. This method involves immunocytochemical (ICC) localization of the Fos and Jun proteins which are rapidly and transiently expressed by neurons in response to physiological stimulation. The localization of the Fos and Jun proteins gives a picture of the pattern of synaptic activity. We will perform ICC for Fos and Jun and computerized morphometric analysis following the ip. or icv injection of IL-1beta. To determine if the activated cells project to CRF neurons, we will use a retrograde tracer to double label the Fos/Jun+ cells. To determine the transmitters used by the Fos/Jun+ neurons of the CNS, we will use double label ICC for Fos and/or Jun and TH, DBH, PNMT, CRF and AVP. 2) The second specific aim will be to localize the distribution of IL-1 receptors on cells of the central CRF system. Using in situ hybridization, we will localize those cells expressing IL-1 receptor mRNA that form the CRF system. Data from this method will then allow further studies on the regulation of the receptor within the CNS. 3) The 3rd specific aim is to determine the effect of IL-1 on the expression of CRF mRNA. These studies will involve both acute and chronic administration of IL-1 and in situ hybridization for CRF mRNA. 4) The final specific aim will be to determine the effect of IL-1 on the blood-brain barrier. Following both acute and chronic IL-1, the integrity of the blood-brain barrier will be determined using iv administration of horseradish peroxidase and localization at both the LM and EM levels. This information may have implications in our understanding of neural-endocrine- immune interactions and in CNS neuropathology.

身体表现出显着维持相对恒定的能力 面对身体或心理的内部环境 需求。 各种自主和内分泌反应，协调 中枢神经系统使身体适应压力。 另外，免疫系统 在面对各种抗原方面起着同等的作用。 越来越多的证据 已经提出，身体的这三个调节系统 持续的沟通。 糖皮质激素是 在免疫反应期间升高，进而抑制免疫反应。 人们认为这种反馈回路始于释放A的免疫细胞 多种细胞因子，包括白介素-1（IL-1）。 IL-1很低 据信具有内分泌学的分子量多肽 神经和免疫功能。 CNS IL-1内已知 导致皮质激素释放因子（CRF）释放到门户 血管，诱发发烧，引起厌食症，增加慢波睡眠，增加 中心自主活动等。虽然前核已经 被视为IL-1在诱导发烧和CRF释放中的作用部位， 通常，我们不知道中枢神经系统内IL-1的作用部位。 我们现在建议研究IL-1相互作用的几个方面 中央CRF神经内分泌系统。 我们将尝试的具体目标 要完成的是：1）确定活化神经元的分布 在用IL-1Beta刺激后的中枢神经系统内。 我们将使用新的 用于在功能上绘制神经元激活的神经解剖技术 IL-1。 该方法涉及免疫细胞化学（ICC）定位 FOS和JUN蛋白迅速且瞬时地表达神经元 响应生理刺激。 FOS的本地化和 JUN蛋白给出了突触活动模式的图片。 我们将 执行ICC用于FOS和JUN和计算机化的形态计算分析 遵循IP。或ICV注入IL-1Beta。 确定是否 激活的细胞投射到CRF神经元，我们将使用逆行示踪剂到 双重标记FOS/JUN+单元。 确定由 CNS的fos/jun+神经元，我们将使用双重标签ICC进行fos和/或 Jun和Th，DBH，PNMT，CRF和AVP。 2）第二个特定目标是 将IL-1受体的分布定位在中央CRF的细胞上 系统。 使用原位杂交，我们将定位这些单元格 表达形成CRF系统的IL-1受体mRNA。 来自此的数据 然后，方法将允许对受体调节的进一步研究 在中枢神经系统内。 3）第三个特定目的是确定IL-1的效果 关于CRF mRNA的表达。 这些研究将涉及急性和 慢性给药IL-1和CRF mRNA的原位杂交。 4） 最终的具体目的是确定IL-1对 血脑屏障。 遵循急性和慢性IL-1，完整性 血脑屏障将使用IV施用确定 LM和EM水平的辣根过氧化物酶和定位。 这 信息可能对我们对神经内分泌的理解有影响 免疫相互作用和CNS神经病理学。