Effect of Nitric Oxide on NGF-Receptor of seminiferous Epithelial Cells

一氧化氮对生精上皮细胞NGF受体的影响

• 批准号: 15591722
• 负责人: ONODA Makoto
• 金额: $ 2.24万
• 依托单位: National Institute of Radiological Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591722/
• 关键词: Seminiferous Epithelial Cell; Sertoli Cell; NGF; NGF-Receptor; p75; TRK; Nitric Oxide; Nitric Oxide Synthase; 精巣上皮細

We investigated the effect of nitric oxide (NO) on neurotrophic factor receptors (NGF-R and TRKs) of rat seminiferous epithelial (Sertoli) cells to elucidate some aspects of the role of NO in intratesticular communication between germ cells and epithelial cells.Immunoreactivity with a specific antibody against NGF gene product localized to germ cells (pachytene spermatocytes and round spermatids) within testis, while gene products of neurotrophic factor receptors such as NGF-R (p75) and Trks (A and B) localized to Sertoli cells. Localization of the NGF-R, TrkA and TrkB in cultured Sertoli cells revealed that the receptors were distributed randomly throughout the cell surface. The diffused immunofluorescence staining represented characteristic immunostaining of plasma membrane receptors that were not pre-clustered in coated pits. In addition, a certain Sertoli cells exhibited that the immunofluorescence staining of these receptors also localized to be in clustered areas of the cells.Exp … More ression of mRNA of these neurotrophic factor receptors was detected by RT-PCR in total RNA samples obtained from the cultured Sertoli cells, and immunoreactive bands of NGF-R (76kDa), TrkA (140kDa) and TrkB (145kDa) were identified in the extracts of cultured Sertoli cells by the Western blot analyses. Expressions of both proteins and mRNAs of the NGF-R, TrkA and TrkB were apparently decreased in the Sertoli cells treated with NOC18 (an NO donor, 0.4mM) for 24h. Treatment with NO donor also resulted in the decline of the NGF-R, TrkA and TrkB at the supranuclear areas of the cells. This indicates that endocytosis of coated pits including the receptors was eliminated by exogenous NO added to the Sertoli cells in the culture. On the other hand, the culture of Sertoli cells with neurotrophic factors such as BDNF (100ng/ml) and NGF (100ng/ml) revealed the up-regulation of Trk (146kDa) detected the Trk-pan antibody, and the combination-treatment of neurotrophic factor and NO donor eliminated the decrease of Trk (146kDa) induced by the NO donor treatment. These results suggest that Sertoli cells express the neurotrophic factor receptors such as NGF-R, TrkA and TrkB and may regulate the proliferation and differentiation of germ cells through the paracrine regulation mechanism within testis. Furthermore, NO produced excessively within testis during acute or chronic pathophysiological conditions perturbs the expression and localization of neurotrophic factor receptors and in turn the signal transduction for spermatogenesis, whereas the ligands of these receptors such as BDNF and NGF might rescue the receptors from the repression induced by the excessive NO and maintain the regulation of normal spermatogenesis. Less

本实验研究了一氧化氮(NO)对大鼠生精上皮(Sertoli)细胞神经营养因子受体(NGF-R和TRKs)的影响，以阐明NO在生殖细胞和上皮细胞间通讯中的作用。针对NGF基因产物的特异性抗体定位于睾丸内的生殖细胞(粗线期精母细胞和圆形精细胞)，而神经营养因子受体的基因产物NGF-R(P75)和Trks(A和B)定位于Sertoli细胞。NGF-R、TrkA和TrkB在培养的Sertoli细胞中的定位表明，受体随机分布于细胞表面。弥散的免疫荧光染色代表了细胞膜受体的特征性免疫染色，这些受体没有预先聚集在被覆的小窝中。此外，某些支持细胞显示，这些受体的免疫荧光染色也定位于细胞的聚集区。Exp…RT-PCR在培养的Sertoli细胞总RNA中检测到这些神经营养因子受体的mRNA表达，Western印迹分析在培养的Sertoli细胞提取物中检测到NGF-R(76 KDa)、TrkA(140 KDa)和TrkB(145 KDa)的免疫反应条带。NOC18(0.4 mM的NO供体)作用于支持细胞24小时后，NGF-R、TrkA和TrkB的蛋白和mRNAs的表达明显减少。没有供体的处理也导致了细胞核上区NGF-R、TrkA和TrkB的减少。这表明，在支持细胞培养中加入外源NO可以消除包被小窝的内吞作用，包括受体。另一方面，支持细胞在神经营养因子BDNF(100 ng/ml)和NGF(100 ng/ml)的共同作用下，Trk(146 KDa)表达上调，检测到Trk-PAn抗体，神经营养因子和NO供体联合处理消除了NO供体处理引起的Trk(146 KDa)的降低。这些结果提示，支持细胞表达神经营养因子受体NGF-R、TrkA和TrkB，可能通过睾丸旁分泌调节机制调节生殖细胞的增殖和分化。此外，在急、慢性病理生理条件下，睾丸内过量产生的NO扰乱了神经营养因子受体的表达和定位，进而干扰了生精信号转导，而这些受体的配体如BDNF和NGF可能会将受体从过量的NO诱导的抑制中解救出来，维持正常的生精调节。较少

项目成果

Radiation-induced deletions in the 5'end region of Notch 1 lead to the formation of truncated proteins and are involved in the development of mouse thymic lymphomas.

辐射诱导的 Notch 1 5 端区域缺失会导致截短蛋白的形成，并参与小鼠胸腺淋巴瘤的发展。

• 发表时间: 2003
• 期刊: Carcinogenesis 24(7)
• 作者: Tsuji;H.;Ishii;H.;H;Ukai.;Katusube;T.;Ogiu;T.
• 通讯作者: T.

Role of nitric oxide in radiation-induced initiation of mammary tumorigenesis in rats

• DOI: 10.1016/s1089-8603(03)00014-4
• 发表时间: 2003-03-01
• 期刊: NITRIC OXIDE-BIOLOGY AND CHEMISTRY
• 影响因子: 3.9
• 作者: Inano, H;Onoda, M
• 通讯作者: Onoda, M

Inano, H., Onoda, M.: "Role of nitric oxide in radiation-induced initiation of mammary tumorigenesis in rats."Nitric Oxide : Biology and Chemistry. 8. 144-148 (2003)

Inano, H., Onoda, M.：“一氧化氮在辐射诱导的大鼠乳腺肿瘤发生中的作用。”一氧化氮：生物学和化学。

Tsuji, H., Ishii, H., Ukai, H., Katsube, T., Ogiu, T.: "Radiation-induced deletions in the 5' end region of Notch 1 lead to the formation of truncated proteins and are involved in the development of mouse thymic lymphomas."Carcinogenesis. 24. 1257-1268 (2

Tsuji, H.、Ishii, H.、Ukai, H.、Katsube, T.、Ogiu, T.：“Notch 1 5端区域的辐射诱导缺失导致截短蛋白的形成，并参与

Nitric oxide produced by inducible nitric oxide synthase is associated with mammary tumorigenesis in irradiated rats

• DOI: 10.1016/j.niox.2004.10.010
• 发表时间: 2005-02-01
• 期刊: NITRIC OXIDE-BIOLOGY AND CHEMISTRY
• 影响因子: 3.9
• 作者: Inano, H;Onoda, M
• 通讯作者: Onoda, M