ARK5 AMP-activated protein kinase and glucose transport

描述(由申请人提供)： 这个项目的直接目标是培训候选人在分子技术(例如，载体的构建，哺乳动物细胞的转基因，和蛋白质转导)方面的培训，以更仔细地研究个别蛋白质在胰岛素和其他刺激刺激葡萄糖运输中的作用，这些刺激是候选人在博士后培训期间研究的。这位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、分离的骨骼肌和活体肌肉。候选人在圣路易斯大学的培训将包括指导实验室研究，出席和参加当地的研究研讨会和国家会议，以及分子生物学的正式课程和负责任的研究行为。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。这种环境为应聘者提供了许多参加研究研讨会和与知名研究人员互动的机会。这位候选人已经证明，AMP激活的蛋白激酶(AMPK)活性的刺激与骨骼肌中胰岛素敏感性的增加有关，他建议研究ARK5，一个新描述的Akt激活的AMPK家族成员，在调节葡萄糖运输中的作用。由于Akt是由肌肉收缩(最近的文献报道和初步数据显示)和胰岛素刺激的，所以这一提议的假设是ARK5在刺激葡萄糖运输的收缩和胰岛素信号通路之间起到串扰作用。候选人的初步数据显示，骨骼肌中含有ARK5。其具体目的是1)确定ARK5是否被肌肉收缩、胰岛素和AMPK激活刺激所激活，2)在肌管中(通过过度表达ARK5、固有活性ARK5的表达和不受Akt调节的ARK5的表达)确定ARK5在无AMPK激活刺激和存在AMPK激活刺激时的胰岛素非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形式转导到肌管中，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人员的过渡，并对候选人为糖尿病研究做出贡献的长期能力至关重要

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