I would like to start a project of inflammatory bowel disease in diabetic mice (diabetes induced by STZ) and I hoped you could help me with the methodology. I know the theory form literature but people use different protocols and practical tips would help a lot.
First, what mice type would you advice BALB/c or black one (age, sex?)
What dose of STZ should I use, how often and for how long?
Should I measure glucose level daily and once a week do the glucose tolerance test? And basically what are the normal glucose levels and when diabetes is diagnosed?
Is the blood collected from a tail or otherwise?
Is abnormal glucose level sufficient to diagnose diabetes or you should as well measure e.g. HbA1c level?
Would you advice to use a reference drug in the diabetes group to create an inner control?
For protocols there is much written on Research Gate. If you look at the answers on my page you will find much advice. The mouse strain does not matter, but the dose of STZ will vary significantly between strains. I would strongly recommend an experiment to titrate the dose in your hands using your protocol as there are many variables to control. Male versus female does not matter. I would pick an age range and stick to it. We tended to use male mice for practical reasons: fewer are needed to maintain the colony. If you are buying in it really does not matter. I think of STZ induced diabetes as a bit like PCR!! There are many protocols which all work. You must work out what works for you being aware of the variables that affect efficiency and toxicity of STZ. I prefer single doses simply because if you spend a bit of time working out the protocol single doses work very well, are reliable and are simpler. With the right dose we see sustained (>100 days) diabetes but if the dose is too low a low incidence of diabetes will be seen and some animals may recover. Blood glucose is good enough for diagnosis and monitoring of diabetes. OGTT is not essential but may be used in some protocols depending what question you are asking. We do not usually use OGTT but for instance in one set of experiments we were titrating the number of islets transplanted and so wanted a more exact readout of transplant function. OGTT was the answer. I have attached a titration experiment for C57Bl6/J mice. We found BALB/c to require a lower dose of STZ. The Y-axis has scrambled but is blood glucose at day 3 post STZ in mmol/l.
Good luck
For protocols there is much written on Research Gate. If you look at the answers on my page you will find much advice. The mouse strain does not matter, but the dose of STZ will vary significantly between strains. I would strongly recommend an experiment to titrate the dose in your hands using your protocol as there are many variables to control. Male versus female does not matter. I would pick an age range and stick to it. We tended to use male mice for practical reasons: fewer are needed to maintain the colony. If you are buying in it really does not matter. I think of STZ induced diabetes as a bit like PCR!! There are many protocols which all work. You must work out what works for you being aware of the variables that affect efficiency and toxicity of STZ. I prefer single doses simply because if you spend a bit of time working out the protocol single doses work very well, are reliable and are simpler. With the right dose we see sustained (>100 days) diabetes but if the dose is too low a low incidence of diabetes will be seen and some animals may recover. Blood glucose is good enough for diagnosis and monitoring of diabetes. OGTT is not essential but may be used in some protocols depending what question you are asking. We do not usually use OGTT but for instance in one set of experiments we were titrating the number of islets transplanted and so wanted a more exact readout of transplant function. OGTT was the answer. I have attached a titration experiment for C57Bl6/J mice. We found BALB/c to require a lower dose of STZ. The Y-axis has scrambled but is blood glucose at day 3 post STZ in mmol/l.
Good luck
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