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

通过收购Seahorse XFe96氧通量分析仪，科尔盖特大学获得了在细胞和细胞器水平上进行多学科研究的奖项。该仪器有助于细胞代谢率的详细分析，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。来自两所主要本科院校（PUI）的不同研究人员都渴望将Seahorse XF分析仪纳入他们的课程和研究项目中。所要求的仪器将在科尔盖特大学和汉密尔顿学院的生物和神经科学的师生合作研究中发挥核心作用，并将促进当前和未来科学领域本科研究培训的卓越性。此外，所要求的海马仪器将对包括神经科学、动物生理学和微生物学在内的课程领域的教学实验室产生重大影响。在本科生研究训练和课程应用之间，该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、病毒感染和癌症等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拓展机会。Seahorse XFe96氧通量分析仪是一种多模态系统，能够测量细胞代谢参数，包括糖酵解速率和线粒体应激。除了在课堂实验室环境中使用外，这项独特的技术将被部署在以学生为中心的研究实验室中，以调查以下研究问题：(1)细胞代谢率如何随狗的体型和年龄而变化，以及它们与整个动物的代谢有何联系？寿命的差异表明，小型犬的寿命明显长于大型犬。因此，狗构成了一个研究生活史和生理学之间联系的信息性群体。我们只能使用海马技术来解开这个基本的生物学规则，因为研究幼狗和老狗的原代成纤维细胞的性质。(2)呼肠孤病毒作为溶瘤剂如何改变细胞代谢途径？呼肠孤病毒是非致病性哺乳动物病毒。然而，关于呼肠孤病毒复制如何改变宿主细胞的代谢状态，我们所知甚少，这是另一个基本的生物学原理，只能用海马XFe96来阐明，因为病毒感染引起的代谢改变只能用完整的细胞来测量。(3)转录因子kr<s:1> ppel样因子4 （KLF4）是否保护线粒体功能异常导致的细胞损伤，以及KLF4是否是细胞周期调节和发育的中介？然而，Klf4在调节细胞代谢途径中的作用仍有待阐明。Seahorse XF技术是测量线粒体代谢和糖酵解的唯一方法（作为替代途径）。(4)当暴露于雌激素样分子DES时，完整秀丽隐杆线虫的全动物细胞表型是如何变化的，DES会降低动物的生育能力？Seahorse XF技术非常适合确定DES对多代细胞代谢的影响。(5)抗坏血酸（AA）对烟草角虫（Manduca sexta）幼虫生长发育有何影响？与饲喂正常aa饲料的幼虫相比，饲喂缺乏aa饲料的幼虫生长更少，发育更慢。昆虫依赖aa生长的机制尚不清楚，海马技术将使我们能够直接检查这种aa依赖过程的代谢基础。总之，这些项目将显著加强科尔盖特大学和汉密尔顿学院的师生研究。

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.