HORMONAL INFLUENCES ON INSECT MOTOR NEURONS DEVELOPING IN CELL CULTURE

激素对细胞培养中昆虫运动神经元发育的影响

• 批准号: 3861647
• 负责人: RICHARD B LEVINE
• 金额: --
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3861647
• 关键词: Lepidoptera; developmental neurobiology; ecdysone; fluorescent dye /probe; hormone regulation /control mechanism; juvenile hormones; metamorphosis; motor neurons; neuroendocrine system; neurotransmitter receptor; tissue /cell culture; voltage /patch clamp; voltage gated channel; Lepidoptera; developmental neurobiology; ecdysone; fluorescent dye /probe; hormone regulation /control mechanism; juvenile hormones; metamorphosis; motor neurons; neuroendocrine system; neurotransmitter receptor; tissue /cell culture; voltage /patch clamp; voltage gated channel

The long-term objectives of this project are to understand the cellular and molecular mechanisms by which steroid and related hormones regulate the development of neuronal circuits. These hormones mediate profound structural and functional changes in the developing central nervous system that are crucial for the maturation of normal behavior. In the proposed study the mechanisms and consequences of hormone action will be investigated in detail by exploiting the advantages of the relatively simple insect nervous system. During insect metamorphosis, identified motor neurons are modified structurally and functionally to accommodate changes in behavior, and several lines of evidence implicate the steroid 20-hydroxyecdysone (20-HE) as a controlling factor. To investigate how ecdysteroids mediate these developmental changes identified motor neurons will be placed in primary cell culture after the incorporation of non-toxic fluorescent labels, which will facilitate cell identification. The ability of identified motor neurons to retain their cell-specific and stage- specific properties in culture will be investigated using a morphometric system to analyze process outgrowth. Intracellular recordings and whole- cell voltage clamp techniques will be used to describe the ligand-gated and voltage-gated currents in neurons isolated from animals of different stages of life. The developmental effects of 20-HE alone, or in combination with other insect hormones. The effects of these hormones on process outgrowth and biophysical properties will be compared for different neurons, and for the same neuron isolated from animals at different stages of development. Finally, the molecular mechanisms of steroid action will be investigated through the addition of protein and mRNA synthesis inhibitors to the culture medium for short periods of time following ecdysteroid exposure. Information obtained by use of this simple system will provide insights into the mechanisms by which steroid hormones and other developmental signals operate in more complex systems.

该项目的长期目标是了解细胞和 类固醇和相关激素调节的分子机制 神经元电路的发展。 这些激素介导了深刻的 发展中枢神经系统的结构和功能变化 对于正常行为的成熟至关重要。 在提议中 研究激素作用的机制和后果将是 通过利用相对的优势来详细调查 简单的昆虫神经系统。 在昆虫变形过程中，发现 运动神经元在结构和功能上经过修改以适应 行为的变化，几条证据暗示了类固醇 20-羟基丁香（20-He）作为控制因素。 调查如何 ecdysteroid介导这些发育变化确定了运动神经元 掺入无毒后，将放置在原代细胞培养物中 荧光标签，这将促进细胞识别。 能力 确定的运动神经元以保留其细胞特异性和分期 将使用形态计量学研究培养的特定特性 分析过程产物的系统。 细胞内记录和全部 细胞电压夹技术将用于描述配体门控和 从不同阶段分离的神经元中的电压门控电流 生活。 独自一人或与之结合的发展效果 其他昆虫激素。 这些激素对过程产物的影响 与不同神经元的生物物理特性和生物物理特性以及 在不同发育阶段从动物中分离出的相同神经元。 最后，将研究类固醇作用的分子机制 通过添加蛋白质和mRNA合成抑制剂 雌激素暴露后短时间内的培养基。 通过使用此简单系统获得的信息将提供见解 进入类固醇激素和其他发育的机制 信号在更复杂的系统中运行。