Dear Miriam,

i would strongly recommend "phenylhydrazine-induced anemia". Treatment of normal RBC with PHZ causes selective association of oxidized alpha-globin chains with the membrane skeleton. PHZ has thus been used to mimic phenotypes of beta-thalassaemia RBC.

Miriam and Greg;

PHZ leaves a lot of junk from damaged RBC that has to be cleaned up by the spleen. some of this is used to produce new RBC. Bleeding of course is a more natural way to induce anemia for mice with normal RBC. Both PHZ and bleeding are dynamic, with loss and recovery, which must be defined in your colony to determine the degree of anemia. As you know, HCT and RBC# can be misleading. The key factor is circulating Hb. Finally, if you need mice with a constant anemia, you might consider one of the genetic anemias due to defects in the cytoskeleton or Fe handling. These reach a steady state of increased RBC production and loss. Let me know if you need more details,

Dear David,

I would indeed prefer to use a genetic model - which model would you recommend ? I would like to use a model which does not interfere with redox signaling or with NO signalling (these are the pathways I would like to investigate in the ischemic tissue). indeed cytoskeleton defects seems to be the best choice. Could you help ? I had a look on the anemia model of the Jackson web site -there are so many. I have a loxP mouse for a gene a would like to make anemic - do you think I may cross them with a genetic model of anemia and then cross them with a specific CRE?

Does your anemia have to be on the B6 background? If not, microcytic might be good as they are reasonably healthy. Their anemia is due to defects in Fe transport. Unfortunately, they may be only available as frozen zygotes and you would have the extra cost to rederive the colony. I suggest phoning the JAX representatives and asking for an expert on the genetic anemias that are not frozen.

Define how anemic you want by the Hb - half normal? Three quarters? And ask which mice are available with such anemias.

I will also ask Luanne Peters, who has done research with mk/mk

thank you so much! I will do it!

I just sent Luanne the questions. Am I correct that since you want to investigate the effects of anemia on the outcome of ischemic events, you just need a constant but reasonably healthy genetic anemia? Is it OK if caused by iron handling defects, or defect in HB proteins, or RBC cytoskeletal proteins? Obviously you will need a non anemic control of the same strain.

yes it is correct!! iron handling defects may interfere with immo-responses or redox state (for example by HO-1) and I would like to avoid it - and if I can cross the mice with my mice (BL6 background) i can use the "anemia gene "negative littermates (by carrying my gene) as a control. Luanne can contact me here or at my university adress. Thank you so much for your help again! Really appreciate it.

Accumulation of alpha particles on the surface of mice blood (in vitro) for short time of exposure cause reduce of blast cells

Miriam - mk/mk anemia is caused by iron handling defects, so no use to you. However Luanne Peters and her colleagues developed a lot of new models with defective hemoglobin protein chains, and also defined models with defective RBC skeletal proteins. Luanne suggests that you just look up in Pub med - Barker JE and Peters LL to find a lot of the papers from 1990s and earlier which provide information on these mice. OF course you still will have to phone the Jackson Production and Sales people to see if they have mice live and breeding, or if they will have to use frozen zygotes which is much much slower and more expensive. Let me know if you have problems.

Thanks again! I will look first the paper then and let u know ( I ordered my CRE line as a frozen embrio and the service was great!)

I found out that Add2 -/- mice and Ank1 -/- mice are for me suitable models. The technical service by jackson also suggested me to contact Dr Peters (who actually he said created one of these two models). I contacted her now per e-mail. Thank you again for your help!

Iron deficient diet can successfully induce iron-deficiency anemia. The diet can be purchased from Research Diet.Inc(0.9mg iron/kg diet). You'd better induce anemia just after weaning (3-4 weeks old), because they are more susceptible to iron deficiency anemia at fast growing age. It only takes 5 weeks, then you will see evident iron deficiency anemia in these mice.

It all depends on what Kind of anaemia you would like to study or use in your mice model. Various genetic anaemias ie haemoglobinopathies , enzymopathies, autoimmune haemolytic anaemias and hypoplastic anaemia has been produced by traditional inbreeding , cross breeding and recent knockout technology. Nutritional deficiencies can be produced by dietary manipulation ir iron deficient diet , folate deficient diet etc. Slow blood loss can produce blood loss related iron deficiency.

Chemical haemolysis can be produced by phenyl hydrazine application systemically.

Trace element deficiency like copper can produce also anaemia. Injection of Benzene can produce MDS and anaemia , ionising radiation and ethyl nitrosourea can produce anaemia as a part of acute leukaemia.

All depends upon what kind of anaemia you want for your study.

If you can use (transient) induced anemia rather than a stable anemia as would be the case for a genetic model, i.p. injection with Phenylhydrazine will do the trick. It's effective, and you can modulate the dose or regimen for more or less anemia as you like.

Here's one paper that described it's use: http://www.sciencedirect.com/science/article/pii/S0925443904000985

are you tested Deferoxamine?Use of iron Chelators in Thalassemia &  Anti Iron poisoning.

There are several ways by which you can create anaemia in mice. The model you chose depends on what you want to study. If you want to study haemolytic anaemia phenylhydrazine is one of the chemicals to use. we used this model to increase reticulocyte count in mice so that we can study the equivalent of Vivax malaria in mice which needs reticulocyte to grow. If you want to study anaemia of marrow suppression Benzene injection ( also cause myelodysplasia )  is one of the techniques. If you want chemotherapy induced suppression of marrow  Busulphan could be used and if you want quick leukaemognesis and anaemia as a consequence there of  ethyl nitrosourea can be used. One who wants to study iron deficiency anaemia chronic blood loss by tail bleeding could be there. Nutritional anaemia also could happen in caged mice through coprophagy or if you overcrowd them  then they eat of part of their legs and paws leading to a combination of bleeding and anaemia of infection . 

We also used foalte deficient diet in caged mice to cause megaloblastic anaemia.

Finally there is a huge number of specially breed or tailor made mice which can mimic haemoglobinopathy , hypoplastic anaemia , autoimmune haemolytic anaemia , myelofibrosis to name a few. These mice could be specially bread or else knock out or knock in mice  . Many examples of such different kind of mice is there eg Belgrade Mice for thalassaemia , SAD mice for Sickle cell disease , w/wv mice or sl/sld mice for different types of hypoplastic anaemia , Newzealand B/W hybrid for immune haemolytic anaemia are some of the examples . In a sinilar way special red cell enzymopenic mice is also available. You have to pick your model depending on your purpose. 

Two commonest cause of anaemia out side acute blood loss is Nutritional ( even with chronic blood loss ) and anaemia of chronic infllamation . Ischaemic events in these two situation which has echoes in human disease isllikely to produce diverse results. eg Nutritional factor ie Iron / B12/ Folate deficiency has far reaching consequences in body economy outside anaemia ( lowered oxygen transport )  same can be said of iron deficiency. Anaemia of chronic inflammation which can be iduced by lipopolysaccharide . carageenan ets primes the immune systems in different ways hence in these sitaution ischaemic events may be compounded by overreaction of our innate and adoptive immune system, In SAD mice sickled redcells produce a combination of ischaemia and anaemia. I feel one model will not serve the purpose. However if one model is to be chosen chronic blood loss may be the simplest model .

Dear, use phenylhydrazine to induce aneamia in the mice and go on with your study.