SULFATED GLYCOLIPID THAT REGULATES DEVELOPMENT OF MALE GERM CELLS

调节雄性生殖细胞发育的硫酸糖脂

• 批准号: 13670138
• 负责人: HONKE Koichi
• 金额: $ 2.3万
• 依托单位: OSAKA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670138/
• 关键词: SPERMATOGENESIS; MEIOSIS; SULFATED GLYCOLIPID; SEMINOLIPID; SULFOTRANSFERASE; KNOCKOUT MOUSE

The sulfated glycolipid, seminolipid occupies more than 90% of total glycolipids in the testis and is specifically expressed in the testis. Seminolipid is biosynthesized in the Golgi membranes of spermatocytes during spermatogenesis. First, a galactosyltransferase called ceramide galactosyltransferase (CGT) acts on the precursor glycerolipid, and synthesizes an intermediate product, galactosylalkylacylglycerol (GaIEAG). Then, a sulfotransferase called cerebroside sulfotransferase (CST) catalyzes the sulfation of GaIEAG, synthesizing the terminal product, seminolipid. We have found that the testes of CST-deficient mice completely lack seminolipid and spermatogenesis is arrested at the metaphase of the first meiosis. These findings indicate that seminolipid is essential for spermatogenesis (Proc.Natl.Acad.Sci.USA 99, 4227-4232, 2002). Next, we have been studying how seminolipid is involved in spermatogenesis. First, we examined the issue whether seminolipid exists in the lipid raft on the plasma membranes of spermatocytes. To this end, we had established the procedure to isolate spermatocytes from mouse testis using a density-gradient centrifugation. As a result, we confirmed that seminolipid is included in the lipid raft. Similarly, we found that another sulfated glycolipid, sulfatide is also included in the lipid raft of brain myelin sheath. Secondly, we have developed a photo-sensitive carbohydrate probe to identify the molecules that interact with seminolipid on the cell-suface of spermatocytes. We are planning to introduce this probe into cultured spermatocytes and detect such molecules in living cells. Thirdly, we have been trying to inject seminolipid into the seminiferous tubules and make the testis to resume spermatogenesis

硫酸化糖脂--胆固醇脂占睾丸中总糖脂的90%以上，并在睾丸中特异性表达。精子发生过程中精母细胞的高尔基体膜合成精脂。首先，称为神经酰胺半乳糖基转移酶（CGT）的半乳糖基转移酶作用于前体甘油脂，并合成中间产物半乳糖基烷基酰基甘油（Galactosylalkylacylglycerol，GaIEAG）。然后，一种称为糖苷磺基转移酶（CST）的磺基转移酶催化GalIEAG的硫酸化，合成最终产物，即磷脂。我们已经发现，CST缺陷小鼠的睾丸完全缺乏磷脂，精子发生在第一次减数分裂的中期被阻止。这些发现表明，胆固醇脂质对于精子发生是必需的（Proc.Natl.Acad.Sci.USA 99，4227-4232，2002）。接下来，我们一直在研究磷脂如何参与精子发生。首先，我们研究了精母细胞质膜上的脂筏中是否存在磷脂的问题。为此，我们建立了一种密度梯度离心法从小鼠睾丸中分离精母细胞的方法。因此，我们证实了脂筏中包含了β-磷脂。同样，我们发现另一种硫酸化糖脂硫苷脂也包含在脑髓鞘的脂筏中。其次，我们研制了一种光敏糖探针，用于鉴定精母细胞表面与磷脂相互作用的分子。我们正计划将这种探针引入培养的精母细胞，并在活细胞中检测此类分子。第三，我们一直在尝试将磷脂注入曲细精管，使睾丸恢复生精功能

项目成果

Saito, T., et al.: "A secreted type of β1,6-N-acetylglucosaminyltransferase V (GnT-V) induces tumor angiogenesis without mediation of glycosylation"J.Biol.Chem.. 277. 17002-17008 (2002)

Saito, T., et al.：“分泌型 β1,6-N-乙酰葡糖胺基转移酶 V (GnT-V) 在不介导糖基化的情况下诱导肿瘤血管生成”J.Biol.Chem.. 277. 17002-17008 (2002)

Ishibashi, T., et al.: "A myelin galactolipid, sulfatide, is essential for maintenance of ion channels on myelinated axon but not essential for initial cluster formation"J.Neurosci.. 22. 6507-6514 (2002)

Ishibashi, T., 等人：“髓磷脂半乳糖脂，硫苷脂，对于维持有髓轴突上的离子通道至关重要，但对于初始簇形成而言不是必需的”J.Neurosci.. 22. 6507-6514 (2002)

Honke, K. et al.: "Paranodal junction formation and spermatogenesis require sulfoglycolipids"Proc.Natl.Acad.Sci.U.S.A.. 99. 4227-4232 (2002)

Honke, K. 等人：“节点旁连接形成和精子发生需要磺基糖脂”Proc.Natl.Acad.Sci.U.S.A.. 99. 4227-4232 (2002)

Ihara, S. et al.: "Prometastatic effect of N-acetylglucosaminyltransferase V is due to modification and stabilization of active matriptase by adding β1-6 GlcNAc branching"J.Biol.Chem.. 277. 16960-16967 (2002)

Ihara, S. 等人：“N-乙酰葡糖胺基转移酶 V 的促转移作用是由于通过添加 β1-6 GlcNAc 分支来修饰和稳定活性基质酶” J.Biol.Chem.. 277. 16960-16967 (2002)

Honke, K., et al.: "Paranodal junction formation and spermatogenesis require sulfoglycolipids"Proc.Natl.Acad.Sci.U.S.A.. 99. 4227-4232 (2002)

Honke, K. 等人：“节点旁连接形成和精子发生需要磺基糖脂”Proc.Natl.Acad.Sci.U.S.A.. 99. 4227-4232 (2002)