CORES

Administrative Mentoring and Recruiting Core

           Drs. Jian-xing Ma and Timothy Lyons will serve as Co-Directors in this Core. The reason to have two Directors in this Core is to combine the basic research experience of Dr. Ma with the clinical skills of Dr. Lyons in diabetes research and patient care. This combination will promote the collaboration between basic scientists and clinical investigators in diabetes research. This combination will also expose the PJI to both basic science training and clinical knowledge and thus, enhance translational research.  This combination will also facilitate the collaboration between the CoBRE with Oklahoma Diabetes Center and GCRC of OUHSC, which are both directed by Dr. Lyons.  A Clinical and Translational Science Award application will be submitted by the GCRC in January 2008 (a planning grant has already been awarded).

            This Core will be responsible for overall administration of the CoBRE including selecting PJIs and mentors, establishing the mentoring program, overseeing the overall budget, organizing meetings of EAC and IAC, organizing seminar series, interacting with all the core directors and PJIs, following the progress and milestones of PJI’s projects and submit progress report to NCRR, and recruiting new investigators to Oklahoma Diabetes Center. This Core will also work closely with the Provost and Dean to recruit new investigators to Oklahoma through the OUHSC Strategic Plan.

Diabetic Animal Core

            The Director of this Core is Dr. Jian-xing Ma.  He has extensive experience with diabetic animal models. Three of the PJI projects will use diabetic animal models. This Core will centralize the induction, monitoring and maintenance of diabetic animals for PJIs. This Core will also coordinate sharing of diabetic animal tissues. The Diabetic Animal Core will provide services to PJIs at no additional charge and thus, reduce the budget of each PJI project.

DESCRIPTION:

            Diabetic rodent models of streptozotocin (STZ)-induced diabetes or genetic diabetes are commonly used in diabetes research. Diabetic animals are associated with high mortality and require intensive cares and characterizations. As most diabetic complications occur after long durations of diabetes, the long-term maintenance and monitoring of diabetic animals are expensive and labor-intensive.

            Dr. Jian-xing Ma has been studying diabetic microvascular complications using a number of diabetic animal models for 15 years. These include both STZ-induced diabetes in mice and rats and genetic diabetic models such as Akita mice, Zuker rats and db/db mice. His group is experienced with induction and monitoring of diabetes using STZ, evaluation of renal functions and retinal vascular changes. In addition, Dr. Ma’s group also has extensive experience with the oxygen-induced retinopathy, a commonly used model for proliferative diabetic retinopathy. To maximize the use of the different tissues (retina, kidney, aorta, heart, liver, serum and urine) of diabetic animals, Dr. Ma’s group has established a collaboration-based Core for sharing diabetic animals with other investigators on campus. His group has been coordinating the sharing of animal tissues with several groups including the labs of Drs. Ming-Hui Zou, Michael Ihnat, Robert Forman, David Kem, Yun Le and John Ash. This collaboration Core has greatly increased the efficiency of diabetic animal research and reduced costs for diabetic animal models in these labs. There is a demand in the OUHSC diabetes research community to expand this Core service to serve all diabetes researchers in Oklahoma.

            The existing diabetes animal Core is located in BSEB313 and has 1500 sq. ft of space. It has facilities and established assays for care and characterization of diabetic mice and rats. These include approved protocols and two chemical hoods for using STZ, facilities for monitoring blood glucose, blood pressure and serum creatinine levels, 12 metabolic cages for mice and 12 for rats available for 24-h urine collection, Oxygen chamber with an oxygen controller for inducing retinal neovascularization, dissecting microscopes, and 3 PCR machines for genotyping of genetic diabetic mice. Two technicians are routinely currently working with diabetic animal models in Dr. Ma’s lab.

SERVICES:

            Three of the PJI projects in this COBRE include experiments using diabetic animal models. The goal of this Diabetic Animal Core is to provide diabetic animals for the PJI projects and to facilitate their projects at no additional costs to the PJI. Dr. Jian-xing Ma will serve as the Director of the Diabetic Animal Core. He has 15-year experience with research using diabetic animal models. The Core director will oversee the entire operation of the Core and is responsible for the interactions with the PJIs and mentors. Dr. Ma will commit one of his technicians who is experienced with diabetic animals to this Core. The Core will recruit a junior faculty member, at Instructor level to be in charge of daily operation of the Core and supervise the work of the technician. This Core will provide the following services.

1. To induce diabetes by STZ injection in rats or mice or in transgenic or gene knockout mice required by the investigators. STZ-induced diabetes is a commonly used type-1 diabetes model. STZ is a compound with a high toxicity and carcinogenicity. Laboratories using STZ will have to go through an Institutional Biosafety Committee (IBC) application process and are required to have safety setup and equipment. All laboratories using STZ are also subjected to regular inspections. The waste with STZ contamination will have to be collected separately, treated and disposed following the IBC guidelines. Apparently, it would be counter-productive if each PJI lab has to obtain a separate approval for using STZ. This core will induce diabetes by injections of STZ for all the PJI projects and mentors of this COBRE. This centralized use of STZ will reduce the time and paper work for these PJIs and also ensure safety. It is also known that STZ dose for induction of diabetes is highly dependent on animal age, sex, body weight, species and strain, and overdose of STZ is associated with animal death. Induction of diabetes by an experienced core will avoid common mistakes and free the PJIs from time-consuming learning process in induction of diabetes.
2. To breed and genotype genetic diabetic mice and rats. Most of the genetic diabetic models do not breed when they are homozygous such as db/db, ob/ob mice and Zuker rats. Homozygous Akita mice do not survive. Heterozygous Akita mice are diabetic and will be bred with wild-type mice. All these genetic diabetic animals will require genotyping. The Core will be responsible for breeding and genotyping of these animals when requested by the PJIs and mentors.
3. To monitor diabetes by measuring hyperglycemia and inject insulin when necessary. The Core will test and record blood glucose levels on a regular basis. When the blood glucose exceeds the desired ranges, insulin will be injected to save the life of the diabetic animal.
4. To collect and record clinical data from diabetic animals. Body weight, urine volume, food and water consumption will be recorded on a regular basis for the diabetic animals.
5. To monitor renal functions of diabetic animals. Urine albumin and creatinine concentrations and serum creatinine levels will be measured on a regular basis or as demanded by individual users to monitor the renal functional damage in diabetic animals.
6. To provide special diet for diabetic animals. Some studies of diabetic animals require maintaining them on a special diet. The Core will be responsible for the dietary control based on the requirement of users.
7. To induce retinal neovascularization in the oxygen-induced retinopathy (OIR) model. OIR mice and rats are commonly used models for retinal neovascularization and vascular leakage. The Core is equipped with a chamber with oxygen controller. The Core will induce retinal neovascularization in mice and rats.
8. To perform specialized assays to evaluate diabetic complications. The Core will perform vascular leakage assay and glomerular filtration rate assay, upon the request from users.
9. Dissect tissues and coordinate the sharing of tissues from diabetic animals. The Core will dissect tissues such as heart, kidney, retina, liver and serum from diabetic animals for the PJI projects. The Core will also establish a tissue bank of diabetic animals and will coordinate sharing of tissues from diabetic animals. This will maximize the use of diabetic animals and reduce costs of diabetic animals.

Histology, Image Acquisition and Image Analysis Core

            The Director of this Core is Dr. James Tomasek, Professor, Dept. of Cell Biology, Dean of Graduate College. He is the Director of an existing Histology Core at OUHSC, Microscopy and Histology Facilities for the Department of Cell Biology, and has extensive experience in fixation, embedding and sectioning of a variety of tissues, photomicroscopy, and immunocytochemistry.  As Director, he will oversee the Core, the Research Program Coordinator, the histology technician, and coordinate the assignment and scheduling of projects from the PJIs and Core participants through the Research Program Coordinator.  He has extensive experience in histological studies and running shared core facilities. Histology is a commonly used technique in this CoBRE. This CoBRE will use and expand existing histology core by covering a full time technician, a portion of the Director and supplies. Using an existing Core Facility is less expensive and more effective, compared to establishing a new Core. This histology Core will serve the PJIs and mentors at no additional fees.

           The purpose of the Histology, Image Acquisition and Image Analysis Core is to provide to the PJIs and diabetes funded investigators, tissue processing, embedding, sectioning, and histochemical staining of mounted slides, quality state of the art instrumentation and expertise to obtain microscopic images with light, epifluorescent, and Nomarski optics; to provide the software for morphometric analysis and to produce publication quality images.  An online signup for image core equipment will be created and made available on the campus net. The module personnel also educate and train PI's, faculty, staff and students in the capabilities and proper use of the sophisticated equipment as well as in tissue processing. The Core comprises 900 sq ft on the 5th floor of the Biomedical Sciences Building and the equipment consists of epifluorescence microscopes with phase and Nomarski differential interference contrast optics interfaced with computer systems for image storage and a computer workstation for image quantitation, and manipulation.  It also contains equipment for processing tissues, embedding, sectioning, cryosectioning and histochemical staining.
 
Services Provided:
 
           Tissue Processing- Because rapid fixation is important for preservation of morphology, the investigators will fix the tissue themselves using fixative and vials provided by the Core.  Depending on the experiment, the investigator will either transfer the tissue to a 70% ethanol solution or give it to the Core histologist in the original fixative.  The samples will be processed in the automated processing center and subsequently manually oriented and paraffin embedded in the embedding center.  The paraffin blocks will be individually trimmed and sectioned by the histologist using the rotary microtome.  The sections will be mounted on slides, stained with H&E or other histochemical stains using the automated slide stainer and coverslipped or left unstained on slides for those investigators using them for immunocytochemical staining.  Some investigators may prefer to have their students and lab personnel cut sections themselves in the interest of time or a specialized requirement of the experiment.  A rotary microtome will be available for sign up and use by these individuals. The Core does not anticipate the instrumentation for automated immunostaining for specific antigens and providing manual immunocytochemical staining as a service is not immediately anticipated.

            For quality control, random sections from an experiment will be routinely viewed under the microscope by the Research Program Coordinator and the Histology Technician prior to giving the slides to the PI.  If appropriate, the investigator will be shown some representative fields for his/her determination as to the experimental value of the sections.  The sections will be mounted and stained with H&E or other histochemical stains and given to the investigator for evaluation.  The investigator will be able to sign up to use the Olympus Provis AX-70 photomicroscope or the Nikon TE2000 inverted epifluorescence microscope to visualize sections using either bright field, Nomarski differential interference contrast, or epifluorscence optics.  Digital images can be captured on either microscope using either the Q-imaging digital camera with Q-capture software or the Hamamatsu Orca AG charge-cooled digital camera with the Openlab suite of imaging software.  Digital images can be analyzed with either Northern Eclipse imaging software, the Openlab suite of imaging software or MetaMorph software at the independent computer workstation.  In all cases the investigator will be asked to save the images directly to a removable hard drive or other media storage device so as not to interfere with the software programs and avoid overloading the hard drive directly connected to the instrument.

           Training- Ms. Carol Haaksma, Research Program Coordinator, will provide training and oversight for the Core microscopes.  An online signup sheet for these microscopes will be available.  Following training, approved individuals will be allowed, after signing up, to use the equipment without supervision.  For members of investigator’s labs who wish to section and stain their own materials, the histology technician will provide training.  For cryostat sections, either members of the investigator’s labs will be trained on the cryostat or the histology technician will cut the sections depending upon the investigator’s needs after discussion with the Director of the Core.

Biostatistics Core

           The Director of this Core is Dr. Chris Aston, Associate Professor, and Director of the GCGC Biostatistics and Bioinformatics Facility. This Core will use and expand the service of the existing GCRC Biostatistics Core by providing additional funds. The Core will assist the PJIs and mentors in experimental design and data analysis. Dr. Aston is a highly experienced biostatistician.

           The goals of the Biostatistics Core are to promote and facilitate the research undertaken by the Junior Investigators in the COBRE, as well as to enhance the research capabilities in the institution.  The Biostatistics Core will provide support for the development of analytical capabilities at all levels of the COBRE, including technicians and mentors, but especially focusing upon the Junior Investigators. As potential collaborators for all research projects, the Biostatistics Core will be an integral part of the research team in the development of research studies; as well as providing ongoing statistical and analytical advice throughout the life of each individual project. Services will be accordingly tailored to the individual Junior Investigator and could include advice or participation in experimental design, database construction, statistics, and publication.

The specific functions include:

1. Providing experimental design advice and training to Junior Investigators (and diabetes researchers at the OUHSC).
2. Assisting, advising, and implementing high quality data management within laboratories.
3. Providing state of the art analysis capabilities for data from, for example, family based genetic epidemiology studies

           Some of the Biostatistics Core’s functions have already been utilized. The Biostatistics Core personnel have already consulted with the PIs regarding experimental design, power/sample size estimations and appropriate statistical analyses for the COBRE projects.