Generation and analysis of mice lacking the prostanoid receptor

缺乏前列腺素受体的小鼠的产生和分析

• 批准号: 07557175
• 负责人: USHIKUBI Fumitaka
• 金额: $ 9.92万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557175/
• 关键词: prostanoid; prostaglandin; thromboxane; prostanoid receptor; knockout mouse; 受容体; 遺伝子ノックアウトマウス; ES細胞; C57BL; 6; プロスタグラジン; トロンボキサンA_2

We constructed the targeting vectors for homologous recombination with each of the eight rypes and subtypes of the prostanoid receptor genes. The vectors were introduced into the ES cells originating from the mouse strain of 129/ola. The blastcystes injected with the ES cells were then intoduced into the uterus of pseudo-pregnant ICR mice, and the chimeric mice were generated. The chimeric mice were crossed with C57BL/6 mice, and the F1 mice were generated followed by F2 mice lacking each of the prostanoid receptor. We have been trying to back-cross each of the mutant mice to C57BL/6, balb/c and DBA1 mice to get the congenic mice.Analyzes of these mice lacking each of the prostanoid receptors have revealed the novel physiological and pathophysiological roles of prstanoids in the body. PGI_2 has roles in increased vascular permiability and transmission of pain sensation in inflammation as well as antithrombotic role. PGF_<2alpha> decreases the plasma progesterone level by inducing the regression of corpus luteum, which then upregulates the expression of the oxytosin receptors in the uterus at term. PGE_2 acts on the EP4 receptor and regulates the function of the ductus arteriosus.These results would contribute to the development of new drugs acting specifically on each prostanoid receptors, and would also present the clues to the application of these drugs.

我们构建了与8种类型和亚型的前列腺素受体基因同源重组的靶向载体。将载体导入129/ola小鼠株的胚胎干细胞中。将注入胚胎干细胞的囊胚植入假妊娠ICR小鼠子宫内，获得嵌合小鼠。将嵌合小鼠与C57BL/6小鼠杂交，产生F1小鼠，随后产生缺乏两种前列腺素受体的F2小鼠。我们一直在尝试将每只突变小鼠与C57BL/6、balb/c和DBA1小鼠进行回交，以获得基因小鼠。对这些缺乏前列腺素受体的小鼠的分析揭示了前列腺素在体内的新的生理和病理生理作用。PGI_2在炎症中具有增加血管通透性和疼痛感觉传递以及抗血栓作用。PGF_<2alpha>通过诱导黄体退行降低血浆孕酮水平，进而上调足月子宫内催产素受体的表达。PGE_2作用于EP4受体，调节动脉导管功能。这些结果将有助于开发特异性作用于每种前列腺素受体的新药，并为这些药物的应用提供线索。

项目成果

Hirata, M., et al.: "Molecular Biology of the Arachidonate Cascade.(分担執筆)" Elsevier Science., 393-404 (1995)

Hirata, M. 等人：“花生四烯酸级联的分子生物学。（贡献者）”Elsevier Science.，393-404 (1995)

Yukihiko Sugimoto: "Failure of parturition in mice lacking the prostaglandin F receptor." Science. 277. 681-683 (1997)

Yukihiko Sugimoto：“缺乏前列腺素 F 受体的小鼠分娩失败。”

Ogawa, Y., et al.: "Structural organization and chromosomal assignment of the human prostacyclin receptor gene." Genomics.27. 142-148 (1995)

Okawa, Y. 等人：“人类前列环素受体基因的结构组织和染色体分配。”

Ushikubi, F., et al.: "Molecular Biology of the Arachidonate Cascade.(分担執筆)" ELSEVIER Science., 343-359 (1995)

Ushikubi, F. 等人：“花生四烯酸级联的分子生物学。（贡献者）” ELSEVIER Science.，343-359 (1995)

Takako Hirata: "Two thromboxane A_2 receptor isoforms in human platelets : opposite coupling to adenylyl cyclase with different sensitivity to Arg^<60> to Leu mutation." Journal of Clinical Investigation. 97. 949-956 (1996)

Takako Hirata：“人血小板中的两种血栓素 A_2 受体亚型：与腺苷酸环化酶相反偶联，对 Arg^<60> 和 Leu 突变具有不同的敏感性。”