Study about alternatively spliced form expression of MIWC water channel in the process of development in rats.

MIWC水通道在大鼠发育过程中选择性剪接形式表达的研究

• 批准号: 08671303
• 负责人: HASEGAWA Hajime
• 金额: $ 1.15万
• 依托单位: Jikei University School of Medicine
• 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-08671303/
• 关键词: aquaporin, MIWC,alternative splicing form, kidney, brain, development; 発生; スプライシングフォーム; 腎臓; 脳; in situ hybridization; MIWC; AQP4; water channel; kidney; brain; development; alternative splicing; gene expression

In last year, we examined changes in expression ratio of alternatively spliced form (SF) of aquaporin 4 (AQP4) vs non-spliced form (NSF) in brain and kidney in the process of development. The expression ratio of both forms was studied by use of quantitative RT-PCR in combination with ELISA method at day 14 and 7 before birth, and day 0,3,7 and 14 after birth in SD male rats. In results, the expression of both SF and NSF of AQP4 were very faint before birth. However, after birth, the expression of SF was clearly detected before enhancement of NSF expression. In this year, we performed in situ hybridization to study the localization of both form of AQP4 in individual developmental period. cRNA probes for SF and NSF of AQP4 were labeled with 35S and applied to frozen tissue sections obtained from individual developing period of rats. The signals corresponding to SF of AQP4 were detected in developing kidney and brain more obviously than NSF signals in rats before birth. This evidence was thought to be conpatible with expression pattern observed in RT-PCR ELISA analysis.These results suggest that alternative splicing of AQP4 gene might play a role as a kind of "switching" to change of AQP4 from inactive form to active form to adjust circumstance.

去年，我们研究了发育过程中大脑和肾脏中水通道蛋白4（AQP4）的可变剪接形式（SF）与非剪接形式（NSF）的表达比的变化。采用定量RT-PCR结合ELISA方法研究SD雄性大鼠出生前14、7天，以及出生后0、3、7、14天两种形式的表达比例。结果显示，AQP4的SF和NSF在出生前的表达都很微弱。然而，出生后，在NSF表达增强之前就清楚地检测到了SF的表达。今年，我们通过原位杂交研究了两种形式的AQP4在个体发育时期的定位。 AQP4 的 SF 和 NSF cRNA 探针用 35S 标记，并应用于从大鼠个体发育期获得的冷冻组织切片。大鼠出生前发育中的肾脏和大脑中检测到的AQP4 SF对应的信号比NSF信号更明显。这一证据被认为与RT-PCR ELISA分析中观察到的表达模式相一致。这些结果表明AQP4基因的选择性剪接可能起到一种“切换”作用，使AQP4从非活性形式转变为活性形式以调节环境。