Molecular biological study of the endolymphatic sac

内淋巴囊的分子生物学研究

• 批准号: 08671969
• 负责人: MORI Nozomu
• 金额: $ 1.47万
• 依托单位: Kagawa Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671969/
• 关键词: endolymphatic sac; hormone receptor; ion channel; PCR; 内リンパ襄

The expression of mRNAs encoding epithelial Na^+ channel (ENaC) and hormone receptors (aldosterone, vasopressin and ANP receptors) in the endolymphatic sac (ES) of the rat was examined by PCR and in situ hybridization. After reverse-transcription of rat ES RNA,cDNA was amplified by PCR using pairs of primers specific to ENaC (-subunit) and aldosterone, vasopressin V1a, V1b, V2, ANP-A and ANP-B receptors. After subcloning of the PCR products, the nucleic acid sequences were determined. Next, the localization of expression of ENaC and those receptors in the ES was examined by in situ hybridization using 35S-labeled cRNA probes.Expressions of EnaC,aldosterone, vasopressin V1a and ANP-A receptors were detected in the ES by PCR.In situ hybridization revealed the presence of aldosterone receptor in epithelial cells of the intermediate portion of the ES,while it failed to show the presence of vasopressin V1a and ANP-A receptors. The results suggest the involvement of aldosterone in Na^+ transport of the ES.

采用PCR和原位杂交技术检测大鼠内淋巴囊（ES）上皮Na^+通道（ENaC）和激素受体（醛固酮、加压素和ANP受体）mrna的表达。将大鼠ES RNA逆转录后，利用ENaC（-亚基）和醛固酮、加压素V1a、V1b、V2、ANP-A和ANP-B受体特异性引物对进行PCR扩增cDNA。对PCR产物进行亚克隆后，确定核酸序列。接下来，使用35s标记的cRNA探针原位杂交检测ENaC和这些受体在ES中的表达定位。PCR检测ES中EnaC、醛固酮、抗利尿激素V1a和ANP-A受体的表达。原位杂交显示ES中间部分上皮细胞中存在醛固酮受体，未发现加压素V1a和ANP-A受体。结果提示醛固酮参与了ES的Na^+转运。

项目成果

Wu, D.: "Outward K+ current in epithelial cells isolated from intermediate portion of endolymphatic sac of guinea pigs." Am.J.Physiol.271. C1765-C1773 (1996)

Wu, D.：“从豚鼠内淋巴囊中间部分分离的上皮细胞中的外向 K 电流。”

Furuta, H.: "Identification of genes expressed in developing rat cochlea by differential display PCR." Proc.Sendai Ear Symposium. 6. 5-6 (1996)

Furuta, H.：“通过差异显示 PCR 鉴定发育中的大鼠耳蜗中表达的基因。”

Mori, N.: "Low-amiloride-affinity Na+ channel in the epithelial cells isolated from the endolymphatic sac of guinea-pigs." Pflugers Arch.Eur.J.Physiol.433. 58-64 (1996)

Mori, N.：“从豚鼠内淋巴囊中分离出的上皮细胞中的低阿米洛利亲和力 Na 通道。”

Furuta,H.: "Expression of mRNA encoding vasopressin Vla,vasopressin V2 and ANP-B receptors in the rat cochlea." Hear.Res.(In press). (1998)

Furuta, H.：“编码加压素 Vla、加压素 V2 和 ANP-B 受体的 mRNA 在大鼠耳蜗中的表达。”

Mori, N.: "Membrane potential in isolated epithelial cells of the endolymphatic sac in the guinea pig." Acta Otolaryngol. (Stockh.). (In press). (1998)

Mori, N.：“豚鼠内淋巴囊分离上皮细胞的膜电位。”