Tamoxifen administration to mice

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(tamoxifen IP)
(pro/con)
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see [http://www.chemicalland21.com/lifescience/phar/TAMOXIFEN%20CITRATE.htm] and [http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=449459 PubChem entry: 4-hydroxytamoxifen]
see [http://www.chemicalland21.com/lifescience/phar/TAMOXIFEN%20CITRATE.htm] and [http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=449459 PubChem entry: 4-hydroxytamoxifen]
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== Tamoxifen administration via chow ==
== Tamoxifen administration via chow ==
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* <font color=green>'''pro''': less labour-intensive and less stressful for the animals</font>
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* <font color=red>'''con''': dosage depends on eating behaviour; induction may take longer then IP</font>
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* tamoxifen formulation in typical mouse food pellets have just become commercially available ([http://www.harlaneurope.com/FFcms/klanten/HAR/index.php?select=3.02.04 Harlan Teklad CRD TAM400/CreER])
* tamoxifen formulation in typical mouse food pellets have just become commercially available ([http://www.harlaneurope.com/FFcms/klanten/HAR/index.php?select=3.02.04 Harlan Teklad CRD TAM400/CreER])
* 400 mg tamoxifen citrate per kg food irradiated pellets
* 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.
* 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.
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* '''advantages''':  less labour-intensive and less stressful for the animals
 
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== Tamoxifen administration via the drinking water ==
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== Tamoxifen administration via drinking water ==
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* <font color=green>'''pro''': less labour-intensive and less stressful for the animals</font>
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* <font color=red>'''con''': dosage limited by low solubility of tamoxifen in water; dosage depends on drinking behaviour; induction may take longer then IP</font>
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#dissolve 100mg tamoxifen in 100µl pure ethanol (10mg/100µl = 100mg/ml tamoxifen)
#dissolve 100mg tamoxifen in 100µl pure ethanol (10mg/100µl = 100mg/ml tamoxifen)
#dilute 100-200x in distilled water to final concentration of 1-0.5mg/ml
#dilute 100-200x in distilled water to final concentration of 1-0.5mg/ml
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compare Gail et al 1999, PMID 10547390, p355: treatment of mice
compare Gail et al 1999, PMID 10547390, p355: treatment of mice
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== Tamoxifen administration via a feeding needle ==
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== Tamoxifen administration using a feeding needle ==
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* <font color=green>'''pro''': dosage more precise than feeding and drinking (less than IP)</font>
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* <font color=red>'''con''': time consuming; stressful for animals</font>
#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.  
#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.  
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== Tamoxifen administration by intraperitoneal injection ==
== Tamoxifen administration by intraperitoneal injection ==
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* '''con''': probably the most stressful method for the mouse; requires proficient handler
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* <font color=green>'''pro''': reliable dosage</font>
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* '''pro''': reliable dosage
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* <font color=red>'''con''': probably the most stressful method for the mouse; requires proficient handler</font>
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== See also ==
== See also ==

Revision as of 11:34, 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.

Contents

Chemical data for tamoxifen

chemical structure of tamoxifen and 4-hydroxy-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

  • pro: less labour-intensive and less stressful for the animals
  • con: dosage depends on eating behaviour; induction may take longer then IP
  • 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.


Tamoxifen administration via drinking water

  • pro: less labour-intensive and less stressful for the animals
  • con: dosage limited by low solubility of tamoxifen in water; dosage depends on drinking behaviour; induction may take longer then IP
  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 using a feeding needle

  • pro: dosage more precise than feeding and drinking (less than IP)
  • con: time consuming; stressful for animals
  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

Tamoxifen administration by intraperitoneal injection

  • pro: reliable dosage
  • con: probably the most stressful method for the mouse; requires proficient handler

See also

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