Studies Of Central Nervous System Functional Anatomy

功能神经解剖学部分结合分子和神经解剖学方法，确定与精神健康、传染病和药物滥用问题有关的神经系统功能的动态方面。我们实验室目前的目标是探索在动物受到应激、炎症刺激或感染时，中枢神经系统(CNS)和免疫系统之间的相互作用。我们的方法是识别免疫挑战诱导的大脑中的细胞和分子成分，并在分子、解剖和功能水平上进一步表征反应。使用组织化学技术绘制了关键的解剖通路和相关的神经递质/受体系统。原位杂交组织化学(ISHH)用于定位和定量神经递质、细胞因子、酶、受体、转录因子和即刻早期基因的mRNA表达，以研究免疫、药物、生理或手术干预的适应性变化。免疫组织化学和双标记技术被用来表征免疫信号分子被诱导表达的细胞的表型。我们已经在大脑中绘制了对细菌内毒素--脂多糖(LPS)急性注射的免疫反应图，2)绘制了可能参与整个大脑传递免疫信号的脑脊液流动路径，3)显示了一旦免疫刺激(LPS)进入血脑屏障后的行为，4)显示了在免疫和非免疫生理挑战条件下神经元和非神经细胞中I类主要组织相容性复合体(MHC)的mRNA激活，5)在癫痫点燃模型中映射了激活素和脑源性神经营养因子(BDNF)的mRNA诱导，以及6)表征了脑室内注射白介素1(IL-1)后导致全脑胶质细胞激活的时间和空间级联事件。被诱导的基因转录本包括IL-1、IL-6、IL-12、肿瘤坏死因子-α(TNF-α)、IL-1受体拮抗剂(IL-1ra)、IL-1转换酶(ICE)、转化生长因子-β(TGF-β)，以及其他免疫信号分子，如抑制因子-kappaB(IkappaB)、诱导型环氧合酶(COX-2)和诱导型一氧化氮合酶(INOS)。在特定的细胞类型(内皮细胞、小胶质细胞、星形胶质细胞和脑膜)和特定的模式(血管、脉络丛和脑室周围器官的高水平表达)中可以诱导mRNAs。目前的工作是研究Toll样受体(TLRs)在检测病原体相关分子和提醒大脑注意外周免疫挑战方面的作用。我们还开始了旨在确定核因子-kappaB转录因子在正常大脑功能和对应激源的反应中所起作用的工作。

The Section on Functional Neuroanatomy combines molecular and neuroanatomical methods to identify dynamic aspects of nervous system function that relate to issues of mental health, infectious disease, and drug abuse. The current objective of our laboratory is to explore the interaction between the central nervous system (CNS) and the immune system in animals that are subjected to stress, inflammatory stimuli, or infections. Our approach is to identify cellular and molecular components in the brain induced by immunological challenges and to further characterize the responses at molecular, anatomical, and functional levels. Key anatomical pathways and relevant neurotransmitter/receptor systems are mapped using histochemical techniques. In situ hybridization histochemistry (ISHH) is used to localize and quantify mRNA expression of neurotransmitters, cytokines, enzymes, receptors, transcription factors, and immediate-early genes in studies of adaptive changes to immunological, pharmacological, physiological, or surgical interventions. Immunohistochemistry and double-label techniques are used to characterize the phenotypes of the cells that show induced mRNA expression of immune signaling molecules. We have 1) mapped in the brain the immune response to acute administration of lipopolysaccharide (LPS), a bacterial endotoxin, 2) mapped the cerebrospinal fluid flow pathways that may be involved in conveying immune signals throughout the brain, 3) shown how an immune stimulus (LPS) behaves once inside the blood-brain barrier, 4) shown activation of class I major histocompatibility complex (MHC) mRNA in both neuronal and non-neuronal cells under conditions of immune and non-immune physiological challenges, 5) mapped activin and brain-derived neurotrophic factor (BDNF) mRNA induction in the kindling model of epilepsy, and 6) characterized the spatial and temporal cascade of events leading to brain-wide glial activation following intracerebroventricular administration of interleukin-1 (IL-1). Gene transcripts shown to be induced include IL-1, IL-6, IL-12, tumor necrosis factor-alpha (TNF-alpha), IL-1 receptor antagonist (IL-1ra), IL-1 converting enzyme (ICE), transforming growth factor beta (TGF-beta), and other immune signaling molecules such as inhibitory factor kappa B (IkappaB), inducible cyclooxygenase (COX-2), and inducible nitric oxide synthase (iNOS). The mRNAs are shown to be induced in specific cell types (endothelia, microglia, astrocytes, and meninges) and in specific patterns (high levels of expression in the blood vessels, choroid plexus and circumventricular organs). Current work examines the role of toll-like receptors (TLRs) in detecting pathogen-associated molecules and alerting the brain about peripheral immune challenges. We are also beginning work aimed at defining the role that the NF-kappaB transcription factor plays in normal brain function and in response to stressors.