Gene expression analysis of febrile neurons expressing prostaglandin receptor EPS

表达前列腺素受体 EPS 的发热神经元的基因表达分析

• 批准号: 13672279
• 负责人: SUGIMOTO Yukihiko
• 金额: $ 2.62万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672279/
• 关键词: preoptic area; vasomotor center; GABA; receptor subtypes; single cell analysis; microarray; pyrogen; aspirin; プロスタランジン; 遺伝子発現プロフィール; シングルセル; RNA増幅; 視床下部; 体温中枢; 発熱

The purpose of this study is (1) to understand what genes are activated upon EP3 signaling in hypothalamic neurons, (2) to investigate signal transduction machanisms exerted by EPS in various cell types, and (3) to clarify which EP subtype is involved in particular pathophysiological actions of PGE2(1) In order to characterize single cell gene expression profiles of hypothalamic EP3-positive neurons, we established and optimized single cell gene expression analysis method. By using this method, we characterized genes changes their expression levels upon PGE2 treatment. PGE2 reduced expression levels of various kinds of genes including GABA A receptors. It should be noted that a GABA agonist has been reported to block PGE2-induced febrile response. We proposed that down-regulation of GABA receptor might be a key event in EP3-expressiong neurons to elicit fever(2) We identified upregulation of specific EP gene expression in immune, ovarian, epidermal, and mucosal tissues upon various kin … More ds of stimuli(3) EP3 has been shown to be coupled to Gi activity, inhibition of adenylate cyclase. We found that EP3 receptors expressed in COS-7 cells showed augmentation of other receptor-stimulated Gs activity in an agonistdependent manner. This EP3 signaling may reflect some aspects of the physiological function of PGE2, such as pain sensation(4) We found that EP2 deficiency attenuated intestinal polyp formation both in number and size in Apc KO mice. We introduced dextransulfate (DSS)induced colitis model into Epdeficient mice. As a result, only EP-4deficient mice showed severe colitis upon 3%DSS treatment thai did not induce significant colitisin wild-type mice. EP4 has been shown to promote epithelial regeneration and to inhibit activation of leukocytes and lymphocytes. We further found that EP3-deficient mice showed increased bleeding tendency and decreased susceptibility to arachidonateinduced thromboembolism compared with wild-type mice. EP3 appears to potentiate TP-induced platelet aggregation by increasing intracellular Ca2+ and/or decreasing cAMP level Less

本研究的目的是（1）了解下丘脑神经元中哪些基因在EP 3信号传导后被激活，（2）研究EPS在各种细胞类型中施加的信号转导机制，以及（3）阐明哪种EP亚型参与PGE 2的特定病理生理作用（1）为了表征下丘脑EP 3阳性神经元的单细胞基因表达谱，建立并优化了单细胞基因表达分析方法。通过使用这种方法，我们表征了PGE 2处理后基因表达水平的变化。PGE 2降低包括GABAA受体在内的多种基因的表达水平。应该注意的是，据报道GABA激动剂可阻断PGE 2诱导的发热反应。我们认为GABA受体的下调可能是EP 3表达神经元引起发热的关键事件。（2）我们发现，在不同程度的发热后，免疫、卵巢、表皮和粘膜组织中特异性EP基因表达上调。 ...更多信息 （3）EP 3与Gi活性偶联，抑制腺苷酸环化酶。我们发现在COS-7细胞中表达的EP 3受体以激动剂依赖的方式增强其他受体刺激的Gs活性。这种EP 3信号传导可能反映了PGE 2生理功能的某些方面，例如疼痛感觉（4）我们发现EP 2缺乏在Apc KO小鼠中在数量和大小上都减弱了肠息肉形成。我们将葡聚糖硫酸酯（DSS）诱导的结肠炎模型引入Ep 3缺陷小鼠。结果，只有EP-4缺陷小鼠在3%DSS治疗后表现出严重的结肠炎，而野生型小鼠则没有诱导明显的结肠炎。EP 4已显示促进上皮再生并抑制白细胞和淋巴细胞的活化。我们进一步发现，与野生型小鼠相比，EP 3缺陷小鼠表现出出血倾向增加和对花生四烯酸诱导的血栓栓塞的易感性降低。EP 3似乎通过增加细胞内Ca 2+和/或降低cAMP水平来增强TP诱导的血小板聚集

项目成果

Kabashima K, Saji T, et al.: "The prostaglandin receptor EP4 suppresses colitis, mucosal damage and CD4 cell activation in the gut"J. Clin. Invest.. 109. 883-893 (2002)

Kabashima K、Saji T 等人：“前列腺素受体 EP4 抑制肠道中的结肠炎、粘膜损伤和 CD4 细胞活化”J.

Fennekohl,A., Sugimoto,Y. et al.: "Contribution of the two Gs-coupled PGE2-receptors EP2-receptor and EP4-receptor to the inhibition by PGE2 of the LPS-induced TNF alphaformation in Kupffer cells from EP2-or EP4-receptor-deficient mice. Pivotal role for t

芬内科尔，A.，杉本，Y.

Sonoshita,M., Takaku,K., Sasaki,N., Sugimoto,Y. et al.: "Acceleration of intestinal polyposis through prostaglandin receptor EP2 in Apc delta 716 knockout mice"Nature Medicine. 7. 1048-1051 (2001)

Sonoshita，M.，Takaku，K.，Sasaki，N.，Sugimoto，Y。

Yoshida, K., Oida, H.et al.: "Stimulation of bone formation and prevention of bone loss by prostaglandin E EP4 receptor"Proc. Natl. Acad. Sci. USA. 99. 4580-4585 (2002)

Yoshida, K., Oida, H.et al.：“前列腺素 E EP4 受体刺激骨形成并预防骨丢失”Proc。

Hatac, N., Yamaoka, K., Sugimoto, et al.: "Augmentation of Receptor-Mediated Adenylyl Cyclase Activity by Gi-Coupled Prostaglandin Receptor Subtype EP3 in a GβγSubunit-Independent Manner"Biochem. Biophys. Res. Commun. 290. 162-168 (2002)

Hatac，N.，Yamaoka，K.，Sugimoto 等人：“Gi 偶联前列腺素受体亚型 EP3 以不依赖 Gβγ 亚基的方式增强受体介导的腺苷酸环化酶活性”Biochem。 162-168 (2002)