Electric Studies of Excitation, Secretion & Contraction

兴奋、分泌的电学研究

• 批准号: 8451325
• 负责人: BERTIL HILLE
• 金额: $ 34.4万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-09-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8451325
• 关键词: 5-Hydroxytryptophan; Address; Adrenergic Receptor; Affective; Attention; Axon; Biochemistry; Biological; Brain; Calcium; Calcium Signaling; Cell Culture Techniques; Cell Membrane Permeability; Cells; Chemical Structure; Cholinergic Receptors; Circadian Rhythms; Complement; Coupled; Cyclic AMP; Cyclic GMP; Cyclic Nucleotides; Diffusion; Ductal Epithelium; Dyes; Electrophysiology (science); Endocrine; Endocrine Glands; Enzymes; Event; Evolution; Excision; Exocytosis; Figs - dietary; Gene Expression; Genes; Grant; High Pressure Liquid Chromatography; Hormones; Indoles; Ion Channel; Islets of Langerhans; Jet Lag Syndrome; Knowledge; Life; Light; Lipid Bilayers; Maintenance; Mammalian Cell; Measures; Melatonin; Membrane Potentials; Messenger RNA; Methods; Microdialysis; Microscope; Molecular Biology; Neurohormones; Neurons; Neurotransmitters; Norepinephrine; Organelles; Pancreatic duct; Performance; Pharmacology; Phospholipase C; Photometry; Photoreceptors; Physiological; Pineal gland; Pinealocyte; Primary Cell Cultures; Property; Rattus; Receptor Activation; Regulation; Relative (related person); Reporting; Rest; Role; Second Messenger Systems; Serotonin; Signal Transduction; Sister; Sleep Disorders; Sleep Wake Cycle; Slice; Stimulus; Structure of superior cervical ganglion; Sympathetic Nervous System; System; Techniques; Testing; Time; Transcript; Tryptophan; Vertebrate Photoreceptors; Vesicle; Work; circadian pacemaker; conditioning; in vivo; operation; patch clamp; receptor; reproductive; response; second messenger; shift work; suprachiasmatic nucleus; trafficking; voltage; voltage clamp

DESCRIPTION (provided by applicant): The pineal gland is an endocrine organ in the brain that is primarily regulated by noradrenaline released by the sympathetic nervous system. It is the part of our circadian clock system that broadcasts the night hormone, melatonin, to all of the body. The night state of pinealocytes is accompanied by extensive changes in gene expression relative to the day state. Many expressed genes are those expressed by a sister lineage, photoreceptors, except that mammalian pinealocytes are not photosensitive. Our overall hypothesis is that pinealocytes switch between two very different electrophysiological excitability states that contribute to regulated secretion of melatonin and to entrainment and maintenance of circadian rhythms. Using biophysical techniques, the night and day states of cultured rat pinealocytes will be contrasted in terms of their ion channel complement, calcium signal dynamics, receptor activation, second messenger levels, and secretory mechanisms. The state changes will be achieved by preincubating cultured primary cells in appropriate neurotransmitters. Cells will be dissociated from rat pineal glands, placed in cell culture, treate with norepinephrine to mimic night, and studied under the microscope with techniques such as whole-cell gigaseal recording, photometry of calcium-sensitive dyes, amperometry and HPLC of neurotransmitters, and use of various live-cell indicators for second messengers. Night pinealocytes are hypothesized to have electrical excitability related to that of photoreceptors, and day pinealocytes are hypothesized to be relatively quiescent. The hypothesis that serotonin is secreted from pinealocytes by quantal exocytosis whereas melatonin and N-acetyl serotonin are secreted by hydrophobic diffusion will also be tested. Understanding the rhythmic secretory mechanisms of the pineal will make an important contribution towards treating sleep disorders, seasonal affective responses to short days, and loss of attention due to jet lag and shift work.

描述（由申请人提供）：松果体是大脑中的内分泌器官，主要由交感神经系统释放的去甲肾上腺素调节。它是我们昼夜节律系统的一部分，它将夜间激素褪黑激素广播到所有身体。松果体的夜间状态伴随着基因表达相对于白天状态的广泛变化。许多表达的基因是由姐妹谱系（感光体）表达的基因，除了哺乳动物的松果体不是光敏的。我们的总体假设是，松果体在两个非常不同的电生理兴奋性之间切换 指出促进褪黑激素分泌以及夹带和维持昼夜节律的指标。使用生物物理技术，培养的大鼠松果体的夜晚状态将与它们的离子通道补体，钙信号动力学，受体激活，第二信使水平和分泌机制形成鲜明对比。状态的变化将通过预孵育适当的神经递质中的原代细胞来实现。细胞将与放置在细胞培养物中的大鼠松腺腺分离，用去甲肾上腺素与模仿夜晚进行处理，并在显微镜下使用诸如全细胞gigaseal记录，钙敏感染料的光度测定，钙敏染料的光度测定，安培剂的光度计和Neurotransters的HPLC的光度计以及对各种现场核素指标的使用和使用。假设夜间松果体具有与光感受器相关的电兴奋性，而白天的松果体被认为是相对静止的。假说5-羟色胺通过量化胞吐作用从松果体分泌，而褪黑激素和N-乙酰血清素被疏水性扩散分泌。了解松果体的节奏分泌机制将为治疗睡眠障碍，短天对季节性情感反应以及由于喷气滞后和转移工作而失去注意力做出重要贡献。