MRI: Acquisition of a Seahorse XFe 96 oxygen flux analyzer for integrative organismal and cellular biology

MRI：购买 Seahorse XFe 96 氧通量分析仪，用于综合有机体和细胞生物学

• 批准号: 1725841
• 负责人: Ana Jimenez
• 金额: $ 27.11万
• 依托单位: Colgate University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1725841&HistoricalAwards=false
• 关键词: MRI; Acquisition; Seahorse; XFe; 96

An award is made to Colgate University to undertake multi-disciplinary research at the cell and organelle level through the acquisition of a Seahorse XFe96 oxygen flux analyzer. This instrument, which facilitates detailed analysis of cellular metabolic rates, will support research in animal physiology, virology, genetics, and neurotransmitter biology. A diverse group of researchers across two primarily undergraduate institutions (PUI) are eager to incorporate the Seahorse XF analyzer within their classes and research programs. The requested instruments will play a central role in collaborative faculty-student research in biology and neuroscience at Colgate University and Hamilton College, and will facilitate current and future excellence in undergraduate research training in the sciences. In addition, the requested Seahorse instruments will have a significant impact on instructional laboratories across the curriculum in areas that include neuroscience, animal physiology, and microbiology. Between undergraduate research training and utilization in courses, this system will impact approximately 150 undergraduates a year. The projects will facilitate research with potential societal benefits in the fields of aging, viral infection, and cancer, and will provide outreach opportunities for high school students interested in STEM fields. The Seahorse XFe96 oxygen flux analyzer is a multimodal system capable measuring cellular metabolic parameters including glycolysis rate and mitochondrial stress. In addition to its use in classroom laboratory settings, this unique technology will be deployed in student-focused research laboratories to investigate the following research questions: (1) How do cellular metabolic rates change with body size and age of dogs and how are they linked to whole-animal metabolism? Differences in longevity demonstrate that small dogs have significantly longer lifespans that larger breed dogs. Thus, dogs constitute an informative group to study connections between life-history and physiology. We can only use Seahorse technology to untangle this basic biological rule due to the nature of working with primary fibroblast cells from young and old dogs. (2) How do reoviruses alter cellular metabolic pathways when used as oncolytic agents? Reoviruses are non-pathogenic mammalian viruses. However, very little is known about how reovirus replication alters a host cell's metabolic state, another basic biological principle that can only be elucidated with the use of a Seahorse XFe96 because metabolic alterations due to viral infections can only be measured using intact cells. (3) Does the transcription factor Krüppel-like factor 4 (KLF4) protect against cellular damage due to malfunctioning mitochondria, and KLF4 is a mediator of cell cycle regulation and development? However, the role of Klf4 in regulating cellular metabolic pathways remains to be elucidated. Seahorse XF technology is the only method to measure mitochondrial metabolism and glycolysis (as an alternative pathway). (4) How does the whole-animal cellular phenotype of intact C. elegans change when exposed to the estrogen-like molecule DES, which decreases fertility in animals? The Seahorse XF technology is perfect to determine the effects of DES on cellular metabolism over multiple generations. (5) What effect does ascorbic acid (AA) have on the growth and development of larvae of the tobacco hornworm, Manduca sexta? Larvae reared on AA-deficient diets grow less and develop more slowly relative to larvae fed a normal AA-containing diet. The mechanisms of AA-dependent growth in insects remain unclear and the seahorse technology will allow us to directly examine the metabolic underpinnings of this AA-dependent process. Together, these projects will significantly enhance student-faculty research at Colgate University and Hamilton College.

高露洁大学被授予通过收购SeaHorse XFe96氧通量分析仪在细胞和细胞器水平进行多学科研究的奖项。该仪器有助于详细分析细胞代谢率，将支持动物生理学、病毒学、遗传学和神经递质生物学的研究。两个主要本科院校(PUI)的不同研究人员团体渴望将SeaHorse XF分析仪纳入他们的课程和研究计划。申请的仪器将在高露洁大学和汉密尔顿学院生物学和神经科学的师生合作研究中发挥核心作用，并将促进目前和未来在科学本科生研究培训方面的卓越表现。此外，所需的海马仪器将对神经科学、动物生理学和微生物学等领域的教学实验室产生重大影响。从本科生科研培训到课程利用，这一系统每年将影响大约150名本科生。这些项目将促进在老龄化、病毒感染和癌症领域具有潜在社会效益的研究，并将为对STEM领域感兴趣的高中生提供外展机会。海马XFe96氧通量分析仪是一种多模式系统，能够测量细胞代谢参数，包括糖酵解率和线粒体应激。除了在课堂实验室环境中使用，这项独特的技术将被部署在以学生为中心的研究实验室，以调查以下研究问题：(1)细胞代谢率如何随狗的身体大小和年龄变化，以及它们如何与整个动物的新陈代谢联系在一起？寿命上的差异表明，小狗比大狗的寿命要长得多。因此，狗构成了一个信息量丰富的群体，用于研究生活史和生理学之间的联系。我们只能使用SeaHorse技术来解开这一基本的生物规则，因为使用来自年轻和老年狗的原代成纤维细胞的性质。(2)呼肠孤病毒作为溶瘤剂时如何改变细胞代谢途径？呼肠孤病毒是一种非致病性哺乳动物病毒。然而，人们对呼肠孤病毒复制如何改变宿主细胞的代谢状态知之甚少，这是另一个基本的生物学原理，只有使用海马XFe96才能阐明，因为病毒感染引起的代谢变化只能使用完整的细胞来测量。(3)转录因子Krüppel-like factor4(KLF4)对线粒体功能障碍引起的细胞损伤有保护作用吗？KLF4是细胞周期调控和发育的中介因子吗？然而，KLF4在调节细胞代谢途径中的作用仍有待阐明。海马XF技术是测量线粒体代谢和糖酵解(作为替代途径)的唯一方法。(4)当接触到雌激素样分子DES时，完整线虫的全动物细胞表型会发生怎样的变化，从而降低动物的生育力？海马XF技术是确定DES对多代细胞新陈代谢的影响的完美选择。(5)抗坏血酸(AA)对烟草天牛幼虫生长发育有何影响？与饲喂含有AA的正常饲料的幼虫相比，在缺乏AA的饲料中饲养的幼虫生长较少，发育较慢。昆虫中依赖AA的生长机制尚不清楚，而海马技术将使我们能够直接检查这种依赖AA的过程的代谢基础。总而言之，这些项目将显著加强高露洁大学和汉密尔顿学院的师生研究。

项目成果

Effects of membrane fatty acid composition on cellular metabolism and oxidative stress in dermal fibroblasts from small and large breed dogs

• DOI: 10.1242/jeb.221804
• 发表时间: 2020-06-01
• 期刊: JOURNAL OF EXPERIMENTAL BIOLOGY
• 影响因子: 2.8
• 作者: Jimenez, Ana Gabriela;Winward, Joshua D.;Champagne, Alex M.
• 通讯作者: Champagne, Alex M.