Difference between revisions of "Tamoxifen administration to mice"

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(Tamoxifen administration via chow)
(See also: links for injection)
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* [http://en.wikipedia.org/wiki/Tamoxifen Tamoxifen in Wikipedia]
* [http://en.wikipedia.org/wiki/Tamoxifen Tamoxifen in Wikipedia]
* [http://ptcl.chem.ox.ac.uk/MSDS/HY/4-hydroxytamoxifen.html material safety 4-hydroxytamoxifen]
* [http://ptcl.chem.ox.ac.uk/MSDS/HY/4-hydroxytamoxifen.html material safety 4-hydroxytamoxifen]
* [http://www.urmc.edu/ucar/manual/drug.htm Guidelines for various methods of fluid administration] by Rochester Medical Center
* [http://www.bu.edu/research/compliance/lacu/lacf/sop/injection-techniques/intraperitoneal.shtml Intraperitoneal
injection into the mouse] by Boston Uni
* [http://www.ahwla.org.uk/site/tutorials/BVA/BVA05-Mouse/Mouse.html Mouse restraining and injection] by AHWLA including a video

Revision as of 08:22, 17 September 2008

back to protocols

One of the uses of tamoxifen in the lab is the activation of enzymes deactivated via an estrogen-binding domain, like Cre-ERT (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains.

Tamoxifen is administered to mice in 2 main ways: via the drinking water or in an oily mix given via a feeding needle. Administration via the drinking water is easier but somewhat less reliable.

Chemical data for tamoxifen

chemical structure of tamoxifen and 4-hydroxy-tamoxifen

Tamoxifen citrate

molecular weight pKa' equilibrium solubility in water at 37°C
563.62 g/mol 8.85 0.5 mg/mL

see [1] and PubChem entry: tamoxifen citrate

4-hydroxytamoxifen (OHT)

Tamoxifen is metabolised to 4-hydroxytamoxifen (often abbreviated OHT).

molecular weight pKa' equilibrium solubility in water at 37°C
563.65 g/mol ? ? mg/mL

see [2] and PubChem entry: 4-hydroxytamoxifen

Tamoxifen administration via chow

  • tamoxifen formulation in typical mouse food pellets have just become commercially available (Harlan Teklad CRD TAM400/CreER)
  • 400 mg tamoxifen citrate per kg food irradiated pellets
  • dosage is based on Kiermeyer et al. 2007 "Optimization of spatiotemporal gene inactivation in mouse heart by oral application of tamoxifen citrate." (PMID 17216603) where efficient induction took 4 weeks. It appears that speed of induction is highly dependent on target genes (LoxP insertion sites) and can be as short as days.
  • advantages: less labour-intensive and less stressful for the animals

Tamoxifen administration via the drinking water

  1. dissolve 100mg tamoxifen in 100µl pure ethanol (10mg/100µl = 100mg/ml tamoxifen)
  2. dilute 100-200x in distilled water to final concentration of 1-0.5mg/ml
  3. use 1% ethanol in distilled water as a negative control
  4. replace water in drinking bottles with tamoxifen solution; let mice drink at will

compare Gail et al 1999, PMID 10547390, p355: treatment of mice

Tamoxifen administration via a feeding needle

  1. 100 mg Tamoxifen-free base (Sigma, T5648) was suspended in 100 μl Ethanol and solved in 900 μl peanut oil (Sigma). This 10 mg/100 μl tamoxifen solution was shaken rigorously at 55°C (Tamoxifen precipitates at RT) and divided in aliquots of 50 μl (1 daily dose = 5mg). Store at -20°C.
  2. Heat to 37°C before administration. Verify that tamoxifen is properly dissolved.
  3. Feed orally to mice with a feeding needle. Fix the mice at their neck and ridge so that the belly is directed to the floor. Hold the mouse tightly so that it can not move the head, but not too tight to avoid choking. Carefully introduce the feeding needle in the mouth behind the tongue (max. 1 cm). It is helpful to mark 1 cm on the needle with an edding pen in order to see how deep you have inserted the needle.
  4. Introduce slowly 50 μl of the Tamoxifen solution with the syringe. Make sure that they swallow (can be seen).
  5. Repeat this procedure for 5 days once a day.

(protocol: Stefan Glaser)

see also Metzger and Chambon, 2001, PMID 11327805, p73

See also

injection into the mouse] by Boston Uni