Basic Protocol 1

Animal Models for Autoimmune and

Inflammatory Disease

BB Rat as Model of Human Insulin-Dependent Diabetes Mellitus

Materials

DP-BB rats <50 days of age (available from Dennis Guberski; see Critical

Parameters and Troubleshooting) Food (e.g., Purina), autoclaved

Drinking water: acidified tap or sterile water, adjusted to pH 3.5 to 4.0 with 1 N HCl

Clidox (Pharmacal Research Laboratories) disinfecting solution: 1 part activator, 1

part base, and 5 parts tap water, prepared fresh Sentinel animals: DR-BB or immunocompetent non-BB rats 0.3-ml heparinized microcuvette capillary tubes (Sarstedt) Glucose reagent strips (Clinistix, Miles Labs)

Microisolator housing (see unit 1.2), autoclaved, and if possible in a specific-pathogen-free (SPF) or viral-antibody-free (VAF) facility Cages, autoclaved Bedding, autoclaved Water bottles, autoclaved 100- to 300-g scale

Calibrated instrumentation for measuring glucose concentration by the glucose oxidase method (e.g., Glucose Analyzer II, Beckman, or One Touch II, Lifescan)

Additional reagents and equipment for management of immunocompromised animals (unit 1.2), serological analysis of blood samples from sentinel rats (see Support Protocol 3), tail vein blood collection (unit 1.7), diagnosis of insulitis (see Support Protocol 1), and treatment of diabetic rats with insulin (see Support Protocol 2)

NOTE: All materials taken into SPF or VAF animal rooms should be vacuum autoclaved, if possible. Nonautoclavable materials and the exposed surfaces of tubes, equipment, and other objects should be disinfected by spraying with Clidox disinfecting solution. Personnel handling the animals should wear sterile gowns or scrub suits, bonnets, masks, gloves, and shoe covers.

Establish animal colony

1. Select and obtain animals <50 days of age of a specific variant of BB rat (see Critical Parameters)

Various colonies of both diabetes-prone (DP-) and diabetes-resistant (DR-) BB rats have been established worldwide. Each is identified according to the location of the production colony—e.g., BB rats from the NIH-sponsored resource colony at the University of Massachusetts Medical School in Worcester are designated as BB/Wor rats and those from Edinburgh are designated as BB/Ed.

2. House the rats in autoclaved microisolators (see unit 1.2), using autoclaved cages, bedding, water bottles, and food in an SPF or VAF facility. Provide the rats with acidified drinking water ad libitum. House sentinel animals in the same animal room, but not in microisolators. Use a laminar flow hood as a workstation for animal husbandry and for experimental procedures. Frequently disinfect animal handling areas, animal racks, and other surfaces with Clidox disinfecting solution to help keep the facility virus free.

An SPF or VAF facility is preferred because the frequency of spontaneous IDDM in both DP- and DR-BB/Wor rats is known to be altered by viral infection. For example, lymphocytic choriomeningitis virus (LCMV; Dyrberg et al., 1988) can prevent diabetes in DP rats, and Kilham rat virus (KRV) can induce IDDM in DR rats (Guberski et al., 1991).

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