Clinical application of seminal plasma sperm motility inhibitor.

精浆精子活力抑制剂的临床应用。

• 批准号: 06454461
• 负责人: IWAMOTO Teruaki
• 金额: $ 4.61万
• 依托单位: St.Marianna University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454461/
• 关键词: Seminal plasma sperm motility inhibitor; Seminal vesicle; Seminal plasma; Asthenozoospermia; Dynein arm; Semenogelin; basic protein; Sperm adhesin family; cDNA; 除膜精子

1. We described the primary structure of SPMI,the coding of boar SPMI cDNA gene, Nucleotide sequence analysis of the 645-bp SPMI cDNA predicts a coded polypeptide of 137 amino acid residues which includes a 21-residue signal peptide and a 116 residues secreted protein. The amino acid sequence of SPMI was found to be highly homologous to AQN-3, a member of spermadhesin family proteins of boar that bind to spermatozoa. Expression of the boar SPMI gene detected by Northern blot analysis revealed that its expression is very abundunt in seminal vesicles and specific to this tissue.2. When SPMI was reacted with normal motile sperm, the motility was inhibited in a dose-dependent manner, and completely suppressed at a SPMI concentration of 1500U/ml. After immediate washing of the immotilized sperm, they resumed movement and recovered 30% motility. However, motility was not recovered at a SPMI concentration of 2000U/ml or more. The mechanism of the inhibition of sperm motility was immunohistochemically examined at electron microscopic level using anti-boar SPMI antibody. SPMI was stained on the surface cell membrane of sperm and not stained on inside cell. SPMI did not penetrate the sperm cell membrane. These findings suggest that sperm can not move physically by the attachment of SPMI to cell membrane or second messenger from the sperm cell membrane attached with SPMI inhibits the activity of dynein ATPase.3. Human SPMI precursor identifies with Semenogelin I,II.We investigated whether gene structure of Semenogelin I,II on infertile patients is different with that on fertile men. Two infertile patients and one fertile man had a deletion of same size, 180bp of Semenogelin I DNA.This deletion of gene was not specific in infetile patients.

1.描述了猪SPMI基因的一级结构，编码猪SPMI基因的645个碱基的核苷酸序列，预测了编码137个氨基酸残基的多肽，包括一个21个残基的信号肽和一个116个残基的分泌蛋白。SPMI的氨基酸序列与AQN-3高度同源，AQN-3是猪精子蛋白家族中与精子结合的蛋白。Northern印迹分析表明，猪SPMI基因在精囊中的表达非常丰富，并且在精囊组织中具有特异性。当SPMI与正常活动精子反应时，精子活力受到抑制，且呈剂量依赖关系，当SPMI浓度为1500U/ml时，精子活力完全被抑制，经立即洗涤后，精子恢复运动，活力恢复30%。然而，当SPMI浓度达到2000U/ml或更高时，其运动能力仍未恢复。用抗猪SPMI抗体在电子显微镜水平上对精子活力抑制的机制进行了免疫组织化学研究。SPMI在精子表面细胞膜上染色，在细胞内不染色。SPMI不能穿透精子细胞膜。这些发现表明精子不能通过SPMI附着在细胞膜上而运动，或者精子细胞膜上的第二信使结合SPMI抑制动力蛋白ATPase的活性。人类SPMI前体与Semenogelin I，II一致，我们研究了不育患者Semenogelin I，II的基因结构是否与生育男性的不同。2例不孕症患者和1例生育男性存在相同大小的Semenogelin I DNA缺失，该缺失在不育患者中不是特异性的。

项目成果

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Shoji Y.，Shimada J，等人：“针对单纯疱疹病毒的反义寡脱氧核苷酸类似物的细胞摄取和生物效应。”

T.Iwamoto Y.Furuichi et al.: "Cloning of boar SPMI gene which is expected specifically in seninal vesicle and codes for a sperm motility inhibitor protein." FEBS Letters. 368. 420-424 (1995)

T.Iwamoto Y.Furuichi 等人：“公猪 SPMI 基因的克隆，预计该基因特异存在于精囊中，并编码精子活力抑制蛋白。”

T.Iwamoto et al.: "Cloning of boar SPMI gene which is expected specifically in seminal vesicle and codes for a sperm motility inhibitor protein." FEBS Letters. 368. 420-424 (1995)

T.Iwamoto 等人：“公猪 SPMI 基因的克隆，预计该基因专门存在于精囊中，并编码精子活力抑制蛋白。”

岩谷若夫、東海林洋子、田村信也、乗松美貫、嶋田甚五郎、水島裕: "抗HSV活性を有するアンチセンス・オリゴDNAの安定性およびウイスル感染細胞における動態の検討。" Drug Delivery System. 11. 427-434 (1996)

Wakao Iwatani、Yoko Tokaibayashi、Shinya Tamura、Minuki Norimatsu、Jingoro Shimada、Yutaka Mizushima：“病毒感染细胞中具有抗 HSV 活性和动力学的反义寡核苷酸稳定性研究。”11. 427 -434（ 1996）

東海林洋子、松原司、大内信香、舟橋恵子、嶋田甚五郎、水島裕: "VEGF mRNAに対するアンチセンスDNAのヒトさい帯静脈の管腔形成抑制効果をカチオン性リポソームが増強。" Drug Delivery System. (in press).

Yoko Tokairin、Tsukasa Matsubara、Nobuka Ouchi、Keiko Funahashi、Jingoro Shimada、Yutaka Mizushima：“阳离子脂质体增强反义 DNA 对 VEGF mRNA 对人脐静脉腔形成的抑制作用”。