Pineal Regulation: Molecular basis of development

松果体调节：发育的分子基础

• 批准号: 7734834
• 负责人: David Klein
• 金额: $ 44.9万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 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/7734834
• 关键词: Address; Adrenergic Agents; Adrenergic Agonists; Adult; Arylalkylamine N-Acetyltransferase; Attention; Binding Sites; Boxing; Brain; Brain region; Cells; Cerebellum; Cervical; Chronobiology Disorders; Circadian Rhythms; Cyclic AMP; Cyclic AMP Response Element; Daily; Development; Disease; Drosophila genus; E-Box Elements; Ectopic Expression; Elements; Embryo; Embryonic Development; Endocrine; Enzymes; Event; Evolution; Excision; Exhibits; Family; Family member; Fetus; Foundations; Ganglionectomy; Gene Cluster; Gene Expression; Gene Expression Profiling; Gene Targeting; Genes; Gland; Glutamate Decarboxylase; Goals; Hand; Heart; Homeobox; Homeobox Genes; Homologous Gene; In Vitro; Introns; Kinesin; Knock-out; Knockout Mice; Knowledge; Learning; Light; Lighting; Localized; Mediating; Melatonin; Messenger RNA; Modeling; Molecular; Monitor; Mutation; N-acetylserotonin; NIH 3T3 Cells; Neonatal; Nerve; Neurons; Norepinephrine; Organism; Pancreas; Pathway interactions; Pattern; Phenotype; Photoreceptors; Phototransduction; Physiological; Pineal gland; Pinealocyte; Play; Polymerase Chain Reaction; Preparation; Production; Proteins; Psyche structure; Quantitative Reverse Transcriptase PCR; RNA Interference; Rattus; Regulation; Regulatory Element; Reporting; Research; Retina; Retinal; Retinal Diseases; Retinal Photoreceptors; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Role; Schedule; Sea Bream; Sequence Analysis; Serotonin; Signal Transduction; Sleep; Staging; Sympathectomy; System; Testing; Time; Tissues; Transcript; Transcriptional Regulation; Transgenes; Transgenic Organisms; Undifferentiated; Variant; Vertebrate Biology; Vertebrate Photoreceptors; Vertebrates; Work; Zebrafish; adrenergic; base; circadian pacemaker; day; driving force; immune function; in vivo; interest; member; mutant; neurodevelopment; postnatal; prevent; promoter; relating to nervous system; trafficking; transcription factor; trend

Evolution of pineal gland: "The defining feature of the pineal gland is the capacity to function as a melatonin factory that operates on a approximately 24 h schedule, reflecting the unique synthetic capacities of the pinealocyte. Melatonin synthesis is typically elevated at night and serves to provide the organism with a signal of nighttime. Melatonin levels can be viewed as hands of the clock. Issues relating to the evolutionary events leading up to the immergence of this system have not received significant attention. When did melatonin synthesis appear in the evolutionary line leading to vertebrates? When did a distinct pineal gland first appear? What were the forces driving this evolutionary trend? As more knowledge has grown about the pinealocyte and the relationship it has to retinal photoreceptors, it has become possible to generate a plausible hypothesis to explain how the pineal gland and the melatonin rhythm evolved. At the heart of the hypothesis is the melatonin rhythm enzyme arylalkylamine N-acetyltransferase (AANAT). The advances supporting the hypothesis will be reviewed here and expanded beyond the original foundation; the hypothesis and its implications will be addressed." From (1) AANAT E-Box: "Arylalkylamine N-acetyltransferase (Aanat) is the penultimate enzyme in the serotonin-N-acetylserotonin-melatonin pathway. It is nearly exclusively expressed in the pineal gland and the retina. A marked rhythm of Aanat gene expression in the rat pineal is mediated by cyclic AMP response elements located in the promoter and first intron. Intron 1 also contains E-box elements, which mediate circadian gene expression in other cells. Here we examined whether these elements contribute to rhythmic Aanat expression in the pineal gland. This was done using transgenic rats carrying Aanat transgenes with mutant E-box elements. Circadian expression of Aanat transgenes was not altered by these mutations. However, these mutations enhanced ectopic expression establishing that the intronic Aanat E-box elements contribute to the gene's pineal specific expression. A similar role of the Aanat E-box has been reported in zebrafish, indicating that Aanat E-box mediated silencing is a conserved feature of vertebrate biology." From (2). "In all ..... species, AANAT activity is regulated at the post-translational level and, to a variable degree, also at the transcriptional level. Here, the transcriptional regulation of pineal aanat (aanat2) of the gilthead seabream (Sparus aurata) was investigated. Real-time polymerase chain reaction quantification of aanat2 mRNA levels in the pineal gland collected throughout the 24-h cycle revealed a rhythmic expression pattern. In cultured pineal glands, the amplitude was reduced, but the daily rhythmic expression pattern was maintained under constant illumination, indicating a circadian clock-controlled regulation of seabream aanat2 In NIH-3T3 cells, the seabream aanat2 promoter was activated by a synergistic action of BMAL/CLOCK and orthodenticle homeobox 5 (OTX5). Promoter sequence analyses revealed the presence of the photoreceptor conserved element and an extended E-box (i.e. the binding sites for BMAL/CLOCK and OTX5 that have been previously associated with pineal-specific and rhythmic gene expression). These results suggest that seabream aanat2 is a clock-controlled gene that is regulated by conserved mechanisms." (From 3) NeuroD: "NeuroD1/BETA2, a member of the bHLH transcription factor family, is known to influence the fate of specific neuronal, endocrine and retinal cells. We report here that NeuroD1 mRNA is highly abundant in the developing and adult rat pineal gland. Pineal expression begins in the 17-day embryo at which time it is also detectable in other brain regions. Expression in the pineal gland increases during the embryonic period and is maintained thereafter at levels equivalent to those found in the cerebellum and retina. In contrast, NeuroD1 mRNA decreases markedly in non-cerebellar brain regions during development. Pineal NeuroD1 levels are similar during the day and night, and do not appear to be influenced by sympathetic neural input. Gene expression analysis of the pineal glands from neonatal NeuroD1 knockout mice identifies 127 transcripts that are down-regulated (>twofold, p < 0.05) and 16 that are up-regulated (>twofold, p < 0.05). According to quantitative RT-PCR, the most dramatically down-regulated gene is kinesin family member 5C ( approximately 100-fold) and the most dramatically up-regulated gene is glutamic acid decarboxylase 1 ( approximately fourfold). Other impacted transcripts encode proteins involved in differentiation, development, signal transduction and trafficking. These findings represent the first step toward elucidating the role of NeuroD1 in the rodent pinealocyte." From (4). Crx/Otx: "Otx2 is a vertebrate homeobox gene, which has been found to be essential for the development of rostral brain regions and appears to play a role in the development of retinal photoreceptor cells and pinealocytes. In this study, the temporal expression pattern of Otx2 was revealed in the rat brain, with special emphasis on the pineal gland throughout late embryonic and postnatal stages. Widespread high expression of Otx2 in the embryonic brain becomes progressively restricted in the adult to the pineal gland. Crx (cone-rod homeobox), a downstream target gene of Otx2, showed a pineal expression pattern similar to that of Otx2, although there was a distinct lag in time of onset. Otx2 protein was identified in pineal extracts and found to be localized in pinealocytes. Total pineal Otx2 mRNA did not show day-night variation, nor was it influenced by removal of the sympathetic input, indicating that the level of Otx2 mRNA appears to be independent of the photoneural input to the gland. Our results are consistent with the view that pineal expression of Otx2 is required for development and we hypothesize that it plays a role in the adult in controlling the expression of the cluster of genes associated with phototransduction and melatonin synthesis." From (5) Pax4: "Pax 4 is a homeobox gene encoding a transcription factor that is essential for embryonic development of the endocrine pancreas. In the pancreas, Pax4 counters the effects of the related Pax6 protein, which is also involved in development of the retina and the pineal gland. In this study, we report that Pax4 is strongly expressed in the pineal gland and retinal photoreceptors of the rat. Pineal and retinal Pax4 mRNA is low in the fetus and increases postnatally; Pax6 exhibits an inverse pattern of expression, being very strongly expressed in both tissues in the fetus. In the adult, the abundance of Pax4 mRNA exhibits a diurnal rhythm in both the pineal gland and retina with maximal levels occurring late in the day. Sympathetic denervation of the pineal gland by superior cervical ganglionectomy prevents the nocturnal decrease in pineal Pax4 mRNA but does not alter the pattern of retinal Pax4 expression. Norepinephrine is released by sympathetic nerves in the pineal gland at night; in the present study, we found that treatment with adrenergic agonists suppresses pineal Pax4 expression in vivo and in vitro. Norepinephrine is known to elevate pineal cyclic AMP; here it was found that treatment with a cyclic AMP mimic reduces pineal Pax4 mRNA. These findings suggest that the nocturnal decrease in pineal Pax4 mRNA is controlled by an adrenergic-cyclic AMP mechanism and that Pax4 may function to mediate adrenergic control of circadian gene expression in the pineal gland." (Rath et al., in preparation). Eya2. Eya2, The homolog of the Drosophila gene eyeless has been identified in the pineal gland and retina. Efforts are underway to characterize the gene products, expression pattern and function.

松果腺的进化：“松果体的定义特征是在大约24小时内运行的褪黑激素工厂的功能，反映了松果体的独特合成能力。褪黑激素合成通常会在夜间升高，并在夜间呈现夜晚蛋白的信号。该系统的浸入量没有引起忧郁症的何时出现在进化的线上该假设的核心是进化。支持该假设的进展将在此处进行审查，并扩展到原始基础之外；该假设及其含义将得到解决。” AANAT E-box：“芳基烷基胺N-乙酰转移酶（AANAT）是5-羟色胺-N-乙酰乙酰乙酸 - 梅拉蛋白途径中的倒数酶。它几乎独立地表达了它在乙腺和视网膜中的触发型源自刺激性的刺激性和刺激器的表达。内含子1还包含E-box元素，在其他细胞中介导了昼夜节律的表达，我们检查了这些元素是否有助于使用AANAT的转基因，这些元素是否会使用AANAT转移。内含子的AANAT电子盒元素有助于基因的松果体特异性表达。来自（2）。 “总共.....种类，在翻译后水平上调节AANAT活性，并且在变化程度上也在转录水平上进行调节。在这里，研究了吉尔特黑德·萨纳特（Sparus aurata）（Sparus aurata）的松果体AANAT（AANAT2）的转录调节。节奏表达模式。启动子序列分析揭示了光感受器保守元件和扩展​​的E-box（即BMAL/Clock和OTX5的结合位点先前与松果体特异性和节奏基因表达相关的结合位点）。这些结果表明，seabream aanat2是一个由保守机制调节的时钟控制基因。”（从3）。 Neurod：“已知BHLH转录因子家族的成员NeuroD1/beta2会影响特定的神经元，内分泌和视网膜细胞的命运。我们在这里报告说，神经轨道1 mRNA在发育中的和成年的小鼠松生腺中高度丰富。在17天的胚胎中，penecter ersection在其他时期的脑部均在其他时期内开始。此后，与小脑和视网膜中发现的水平相当，在发育过程中，神经1 mRNA在非脑脑区域中显着降低。下调（>双重，p <0.05）和16个上调的（>两倍，p <0.05），根据定量的RT-PCR，最急剧下调的基因是驱动蛋白家族成员5C（约100倍），大约是最显着上调的基因和牙孔酸胶酶1（近似）。其他受影响的转录本编码涉及分化，发育，信号转导和运输的蛋白质。这些发现代表了阐明NeuroD1在啮齿动物松果体中的作用的第一步。 Crx/Otx: "Otx2 is a vertebrate homeobox gene, which has been found to be essential for the development of rostral brain regions and appears to play a role in the development of retinal photoreceptor cells and pinealocytes. In this study, the temporal expression pattern of Otx2 was revealed in the rat brain, with special emphasis on the pineal gland throughout late embryonic and postnatal stages. Widespread high OTX2在胚胎大脑中的表达在成年人中逐渐限制为松果体（圆锥体型同源物）。没有显示昼夜变化，也没有受到移除交感神经输入的影响，表明OTX2 mRNA的水平似乎与腺体的光负输入无关。我们的结果与以下观点相一致：开发需要松果体表达OTX2，我们假设它在成年人中起作用在控制与光转移和褪黑激素合成相关的基因簇的表达中起作用。 Pax4: "Pax 4 is a homeobox gene encoding a transcription factor that is essential for embryonic development of the endocrine pancreas. In the pancreas, Pax4 counters the effects of the related Pax6 protein, which is also involved in development of the retina and the pineal gland. In this study, we report that Pax4 is strongly expressed in the pineal gland and retinal photoreceptors of the rat. Pineal and retinal PAX4 mRNA在胎儿中较低，PAX6表达的表达方式很大，在成年人的两个组织中都非常表达。松果体PAX4 mRNA的夜间降低，但不会改变视网膜PAX4表达的模式。在本研究中，我们发现肾上腺素能激动剂的治疗抑制了体内和体外的松果体PAX4表达。已知去甲肾上腺素可以提升松果体环状AMP；在这里发现，用环状AMP模拟的处理可减少松果体PAX4 mRNA。这些发现表明，松果体PAX4 mRNA的夜间降低受肾上腺素能 - 循​​环AMP机制的控制，并且PAX4可能起作用可介导松果体中昼夜节律基因表达的肾上腺素能控制。”（Rath等人在制备中）。 Eya2。 Eya2，果蝇基因无眼的同源物已在松果体和视网膜中鉴定出来。 正在努力表征基因产物，表达模式和功能。

项目成果

NeuroD1: developmental expression and regulated genes in the rodent pineal gland.

NeuroD1：啮齿动物松果体中的发育表达和调节基因。

• DOI: 10.1111/j.1471-4159.2007.04605.x
• 发表时间: 2007
• 期刊: Journal of neurochemistry
• 影响因子: 4.7
• 作者: Munoz,EstelaM;Bailey,MichaelJ;Rath,MartinF;Shi,Qiong;Morin,Fabrice;Coon,StevenL;Moller,Morten;Klein,DavidC
• 通讯作者: Klein,DavidC

Expression of the Otx2 homeobox gene in the developing mammalian brain: embryonic and adult expression in the pineal gland.

Otx2 同源盒基因在发育中的哺乳动物大脑中的表达：胚胎和成人在松果体中的表达。

• DOI: 10.1111/j.1471-4159.2006.03773.x
• 发表时间: 2006
• 期刊: Journal of neurochemistry
• 影响因子: 4.7
• 作者: Rath,MartinF;Munoz,Estela;Ganguly,Surajit;Morin,Fabrice;Shi,Qiong;Klein,DavidC;Moller,Morten
• 通讯作者: Moller,Morten