Metabolic and Cellular Physiology core facility, developed as part of COBRE ACCORD, provides technical expertise, equipment, and personnel to ACCORD COBRE and Marshall University investigators who wish to work with obesity and related metabolic disorders such as diabetes, hyperlipidemia, cardiovascular diseases etc. The primary goals of the core are: (1) to generate in vivo models of obesity, ex vivo and in vitro cellular systems in a time-sensitive and cost-effective manner; (2) to provide expertise on comprehensive data analysis of metabolic parameters and cellular factors based on the investigator’s research project; (3) to foster interactions and synergy among COBRE ACCORD investigators, pilot grant awardees and other Marshall University investigators with similar research interests.
Following is the list of services that are provided by the core facility:
(1) Generate in vivo models of obesity and related core services: The core generates various genetic, or diet-induced obesity animal models (rat and mouse models). In addition to generating the animal models of obesity, the core also provides the following services to study the effect of obesity/specific diet on the metabolic parameters of the animal system. (1A). Calorimetric data acquisition: The core provides a range of services on indirect assessment of calorimetric parameters including oxygen consumption (VO2), carbon dioxide production (VCO2), respiratory exchange ratios (RER), food and water consumption, activity/movement levels, core body temperature and heart rate, using Oxyletpro system (Harvard apparatus) for both mouse and rat models of obesity. The Oxyletpro system also helps to collect urine and feces from the experimental animals to determine desired metabolic parameters. (1B). DSI implantable telemetry devices: The core helps to generate continuous real-time telemetric data (blood pressure and blood glucose levels) from the rat and mouse models of obesity. The small implanted rodent devices provide reliable and accurate data from the animal models through the entire period of the study. The core also houses a centralized instrumentation facility that will include a variety of analytical instruments to support obesity research. Following are some of the instruments that are included currently in the core facility to measure metabolic and cellular factors from a biological specimen: Nova Prime plus analyzer, IDEXX VetTest and SpectraMax i3x Multi-Mode Microplate Reader.
Using the centralized facility to generate animal models of obesity provides the investigators the assurance of a high degree of quality control, procedures, protocols and reproducibility of data. Additionally, it enables the synergizing of research data obtained from various organ systems of the same animal, thus facilitating a holistic understanding of obesity and its metabolic disorders. Finally, the facility enables the maximum use of available animal and research resources integrated with cost-effective approaches.
(2) Ex vivo organ culture facility for human and mouse 3D organoid/enteroid model systems: The goal of this facility is to offer expertise, intellectual and material support to COBRE and Marshall University investigators who would like to generate and use 3D human or mouse organoid model systems for their research.
(3) Gene knockdown and gene rescue experiments with functional proteomics: This facility provides technical expertise and assistance with designing and performing shRNA/siRNA and gene rescue experiments. In addition, to provide signaling insights in relation to the gene knockdown and rescue experiments, the core also provides antibody-based functional proteomics. The core offers technical assistance on protein extraction from cells or tissues, performs the antibody-based array, and also performs primary data analysis using statistical software to determine signaling intensity, relative protein expression and modification.
(4) Imaging core facility: Headed by Dr. Norton, the imaging core facility serves as a sub-core under MCP core in COBRE ACCORD program. The imaging core provides the instrumentation infrastructure and learning environment required to support the research of COBRE investigators and COBRE pilot grant awardees.