Characterization of the epididymis-specific lipocalin promoters to develop male contraceptive drugs in future

附睾特异性脂质运载蛋白启动子的表征，用于未来开发男性避孕药物

• 批准号: 18591792
• 负责人: SUZUKI Kichiya
• 金额: $ 2.53万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591792/
• 关键词: epididymis; gene regulation; androgen receptor; sperm maturation; male reproduction; 精単上体; プロモーター解析

Sperm maturation process is one of the important function of the epididymis. If the process is specifically modulated, the sperm function for fertilizing ability may be disturbed. Therefore we hypothesized that characterization of epididymis-specific promoters would lead to the innovation of the male contraceptive drugs in future. As an initial study, we investigated the gene regulation mechanism of the Lipocalin 5 gene promoter. Lipocalin 5 is synthesized and secreted by the principal cells of the mouse middle/distal caput epididymidis and its regulation is androgen-dependent. We determined important cis-regulatory element between the 1.2 and 1.3 kb upstream region from transcription starting site using transgenic mice and epididymal cells. We also discovered that two androgen receptor binding sites and Foxal binding site exist in the 100 by fragment and that these molecules regulate gene expression of the Lipocalin 5 gene (Yu X, Suzuki K et al. Mol EndocrinoL 2007). Furthermore, we studied the Lipocalin 8 gene promoter as a second model, which is expressed in the initial segment and is regulated by transcription factors. We used same strategy as used for the Lipocalin 5 promoter study. Using transgenic mice, we established three lines carrying mutated Lipocalin 8 promoters ligated to the CAT reporter gene (3.0 kb, 1.7 kb, 0:3 kb). We confirmed that reporter gene expression level is decreased between 3.0 kb and 1.7 kb indicating that the 1.3-kb fragment contains important cis-regulatory element for the epididymis-specific gene expression, especially for the initial segment region.

精子的成熟过程是附睾的重要功能之一。如果这一过程被特别调节，精子的可交配性功能可能会受到干扰。因此，我们推测，附睾特异性启动子的特性将导致未来的男性避孕药物的创新。作为初步研究，我们研究了Lipocalin 5基因启动子的基因调控机制。脂质运载蛋白5由小鼠附睾中/远端头的主要细胞合成和分泌，其调节是雄激素依赖性的。我们确定了重要的顺式调控元件之间的1.2和1.3 kb的上游区域从转录起始位点使用转基因小鼠和附睾细胞。我们还发现，在100 bp片段中存在两个雄激素受体结合位点和Foxal结合位点，并且这些分子调节脂质运载蛋白5基因的基因表达（Yu X，Suzuki K等人，Mol Endocrinol 2007）。此外，我们研究了Lipocalin 8基因启动子作为第二个模型，它在起始片段中表达，并受转录因子的调控。我们使用与用于脂质运载蛋白5启动子研究相同的策略。使用转基因小鼠，我们建立了三个系，其携带突变的Lipocalin 8启动子，连接到CAT报告基因（3.0 kb，1.7 kb，0：3 kb）。我们证实，报告基因的表达水平在3.0 kb和1.7 kb之间下降，表明1.3 kb片段包含重要的附睾特异性基因表达的顺式调控元件，特别是在起始片段区域。

项目成果

Positive expression of the immunoglobulin superfamily pretein IZUMO on human sperm of severe male infertile patients.

免疫球蛋白超家族蛋白IZUMO在严重男性不育患者精子中的阳性表达。

• 发表时间: 2007
• 期刊: Fertil. Steril. (In press)
• 作者: Hayasaka S;Terada Y. et al.
• 通讯作者: Terada Y. et al.

Epididymis-Specific Lipocalin Promoter

附睾特异性脂质运载蛋白启动子

• 发表时间: 2006
• 作者: 小橋元;他;鈴木吉也;Invited speaker
• 通讯作者: Invited speaker

Epididymis-specific promoter driven gene targeting

附睾特异性启动子驱动的基因靶向

• 发表时间: 2006
• 作者: Invited Speaker;Kichiya Suzuki
• 通讯作者: Kichiya Suzuki

Management of the pain associated with endometriosis : an update of the painful problems (review)

子宫内膜异位症相关疼痛的管理：疼痛问题的更新（综述）

• 发表时间: 2006
• 期刊: Tohoku J Exp Med 210
• 作者: Ozawa Y.;Murakami T.et al.
• 通讯作者: Murakami T.et al.

Lactation and risk of endometrial cancer in Japan: a case-control study.

• DOI: 10.1620/tjem.208.109
• 发表时间: 2006-01
• 期刊: The Tohoku journal of experimental medicine
• 作者: C. Okamura;Y. Tsubono;K. Ito;H. Niikura;T. Takano;S. Nagase;K. Yoshinaga;Y. Terada;T. Murakami-T.-Mu