THE DYNAMIC CELLULAR LOCALIZATION OF TRPM1

TRPM1 的动态细胞定位

• 批准号: 7960156
• 负责人: Elena Oancea
• 金额: $ 4.13万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-04 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960156
• 关键词: Behavioral Research; Cell membrane; Cell physiology; Computer Retrieval of Information on Scientific Projects Database; Diagnostic; Electron Microscopy; Family; Fluorescence; Funding; Genes; Goals; Grant; Human; Image; Incidence; Institution; Ion Channel; Lead; Melanoma Cell; Molecular; Mus; Neoplasm Metastasis; Property; Research; Research Personnel; Resources; Role; Skin Cancer; Source; Subcellular structure; System; Tumor Suppressor Proteins; United States National Institutes of Health; Viral; cancer cell; interest; melanocyte; melanoma; member; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Malignant melanoma is the deadliest form of skin cancer that continues to increase in incidence at a rate of 3% per year. Melastatin (TRPM1) was discovered as a gene down-regulated in highly metastatic tumors in mice and humans, suggesting that TRPM1 expression may be an indicator of melanoma aggressiveness. This finding raised the interesting possibility that TrpM1 might be a tumor suppressor, an unprecedented function for an ion channel. Despite being the founding member of the TrpM family of ion channels, very little is known about the molecular properties and cellular functions of TrpM1. The goal of the proposed research is understanding the function of TrpM1 and the connection between TrpM1 and melanoma. Determining the molecular mechanisms by which TrpM1 is involved in normal and malignant cells could lead to significant progress in the diagnostic and treatment of melanoma. To understand the role of TrpM1 in cellular function, it is critical to characterize its sub-cellular localization and cellular dynamic. To express fluorescently tagged TRPM1 in primary human melanocytes and melanoma cells we will use a lenti-viral system. We will then employ confocal imaging, total internal reflection fluorescence (TIRF) and electron microscopy (EM) to determine the intracellular versus plasma membrane distribution of TrpM1, the dynamic properties of the intracellular structures containing TRPM1 and their identity. The results of the proposed experiments will represent a significant step towards understanding the function of TrpM1 in melanocytes.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 恶性黑色素瘤是最致命的皮肤癌，发病率以每年3%的速度持续增加。Melastatin(TRPM1)基因在小鼠和人类的高转移性肿瘤中被发现下调，这表明TRPM1的表达可能是黑色素瘤侵袭性的指标。这一发现提出了一个有趣的可能性，即TrpM1可能是一种肿瘤抑制因子，对于离子通道来说，这是一种前所未有的功能。 尽管是TrpM离子通道家族的创始成员，但人们对TrpM1的分子特性和细胞功能知之甚少。这项拟议研究的目标是了解TrpM1的功能以及TrpM1与黑色素瘤之间的联系。确定TrpM1参与正常和恶性细胞的分子机制可能会在黑色素瘤的诊断和治疗方面取得重大进展。 要了解TrpM1在细胞功能中的作用，关键是要表征它的亚细胞定位和细胞动力学。为了在原代人类黑素细胞和黑色素瘤细胞中表达荧光标记的TRPM1，我们将使用Lenti病毒系统。然后，我们将使用共聚焦成像、全内反射荧光(TIRF)和电子显微镜(EM)来确定TrpM1在细胞内和质膜上的分布，以及含有TRPM1的细胞内结构的动态特性和它们的特性。 拟议的实验结果将代表着朝着了解TrpM1在黑素细胞中的功能迈出的重要一步。