ARK5 AMP-activated protein kinase and glucose transport

描述（由申请人提供）： 该项目的直接目标是对候选人进行分子技术（例如载体构建、哺乳动物细胞转染和蛋白质转导）方面的培训，以更仔细地检查候选人在博士后培训期间研究的胰岛素和其他刺激物刺激葡萄糖转运中各个蛋白质的作用。该候选人的职业目标是将糖尿病研究应用于广泛的模型，包括培养的肌管、分离的骨骼肌和体内肌肉。候选人在圣路易斯大学的培训将包括指导实验室研究、出席和参加当地研究研讨会和全国会议，以及分子生物学和负责任的研究行为的正式课程。邻近机构华盛顿大学的糖尿病研究人员将提供额外的技术培训、职业指导和研究咨询。该环境为候选人提供了大量参加研究研讨会并与知名研究人员互动的机会。该候选人已证明，刺激 AMP 激活蛋白激酶 (AMPK) 活性与骨骼肌胰岛素敏感性增加有关，他提议研究 ARK5（一种新描述的 Akt 激活 AMPK 家族成员）在调节葡萄糖转运中的作用。由于 Akt 受到肌肉收缩（如最近文献报道和初步数据所示）和胰岛素的刺激，因此该提议的假设是 ARK5 在刺激葡萄糖转运过程中有助于收缩信号通路和胰岛素信号通路之间的串扰。候选人的初步数据显示骨骼肌含有ARK5。具体目标是 1) 确定 ARK5 是否被肌肉收缩、胰岛素和 AMPK 激活刺激激活，2) 确定肌管中（通过 ARK5 的过表达、组成型活性 ARK5 的表达以及不能被 Akt 调节的 ARK5 的表达）在存在和不存在 AMPK 激活刺激的情况下 ARK5 在胰岛素依赖性葡萄糖转运和胰岛素敏感性中的作用，以及3)通过将ARK5形式转导至肌管，确定ARK5在AMPK激活刺激诱导的胰岛素敏感性中的急性作用。拟议的培训将培养候选人向独立研究员的过渡，对于候选人长期为糖尿病研究做出贡献的能力至关重要

DESCRIPTION (provided by applicant): The immediate goal of this project is to train the candidate in molecular techniques (e.g. construction of vectors, transfection of mammalian cells, and protein transduction) to more closely examine the roles of individual proteins in the stimulation of glucose transport by insulin and other stimuli that the candidate studied during his postdoctoral training. The candidate's career aim is to apply diabetes research to a wide range of models, including cultured myotubes, isolated skeletal muscle, and muscle in vivo. The candidate's training at Saint Louis University will include mentored laboratory research, attendance of and participation in local research seminars and national conferences, and formal coursework in molecular biology and the responsible conduct of research. Diabetes researchers at a neighboring institution, Washington University, will provide additional technical training, career guidance, and research consultation. The environment offers numerous opportunities for the candidate to attend research seminars and interact with established researchers. The candidate has shown that stimulation of AMP-activated protein kinase (AMPK) activity is associated with increased insulin sensitivity in skeletal muscle, and he proposes to examine the role of ARK5, a newly-described Akt-activated AMPK family member, in the regulation of glucose transport. Since Akt is stimulated by muscle contractions (as recently reported in the literature and shown in preliminary data) and insulin, the hypothesis for this proposal is that ARK5 contributes to cross-talk between contraction- and insulin- signaling pathways in the stimulation of glucose transport. The candidate's preliminary data shows that skeletal muscle contains ARK5. The specific aims are 1) to determine whether ARK5 is activated by muscle contractions, insulin, and AMPK-activating stimuli, 2) to determine in myotubes (by means of overexpression of ARK5, expression of a constitutively active ARK5, and expression of ARK5 that cannot be regulated by Akt) the role of ARK5 in insulin-independent glucose transport and insulin sensitivity in the absence and presence of AMPK-activating stimuli, and 3) to determine, by transduction of ARK5 forms into myotubes, the acute role of ARK5 in the insulin sensitivity induced by AMPK-activating stimuli. The proposed training will develop the candidate in his transition to an independent researcher and is critical to the candidate's long-term ability to contribute to diabetes research