Tamoxifen administration to mice - Revision history

Jakob Suckale: /* Tamoxifen administration using a feeding needle */

2010-01-20T15:09:53Z

Tamoxifen administration using a feeding needle

←Older revision		Revision as of 15:09, 20 January 2010
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	* <font color=red>'''con''': time consuming; stressful for animals</font>		* <font color=red>'''con''': time consuming; stressful for animals</font>

		+	final concentration: 100µg/µl ~ '''1mg/10µl'''
	#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.
-	#~~Heat~~ to 37°C before administration. Verify that tamoxifen is properly dissolved.	+	#heat to 37°C before administration. Verify that tamoxifen is properly dissolved.
-	#~~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.	+	#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.
-	#~~Introduce~~ slowly 50 μl of the ~~Tamoxifen~~ solution with the syringe~~. Make sure that they swallow (can be seen).~~	+	#slowly introduce 50 μl of the tamoxifen solution with the syringe; assure swallowing;
-	#~~Repeat~~ this procedure for 5 days once a day.	+	#e.g. repeat this procedure for 5 days once a day (efficiency of induction depends on Cre and transgene; determine your own dosage)

	(protocol: Stefan Glaser)		(protocol: Stefan Glaser)

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

	== Tamoxifen administration by intraperitoneal injection ==		== Tamoxifen administration by intraperitoneal injection ==

Jakob Suckale at 13:03, 7 April 2009

2009-04-07T13:03:31Z

←Older revision		Revision as of 13:03, 7 April 2009
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	One of the uses of [[tamoxifen]] in the lab is the activation of enzymes deactivated via an estrogen-binding domain, like Cre-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains. There are several ways to administer tamoxifen to mice each with advantages and disadvantages. If precise dosage is not paramount, administration via food is the gentlest to the animal. Keep in mind that induction varies depending on Cre and target genes.		One of the uses of [[tamoxifen]] in the lab is the activation of enzymes deactivated via an estrogen-binding domain, like Cre-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains. There are several ways to administer tamoxifen to mice each with advantages and disadvantages. If precise dosage is not paramount, administration via food is the gentlest to the animal. Keep in mind that induction varies depending on Cre and target genes.
-

	== Tamoxifen administration via chow ==		== Tamoxifen administration via chow ==

Jakob Suckale: /* See also */

2009-04-07T13:02:47Z

Jakob Suckale: illustration

2009-04-07T12:35:40Z

illustration

←Older revision		Revision as of 12:35, 7 April 2009
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	{{back to protocols}}		{{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-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains. There are several ways to administer tamoxifen to mice each with advantages and disadvantages. If precise dosage is not paramount, administration via food is the gentlest to the animal. Keep in mind that induction varies depending on Cre and target genes.
-
-	~~== Chemical data for [[tamoxifen]] ==~~

	[[Image:Tamoxifen(OH).gif\|right\|thumb\|150px\|chemical structure of tamoxifen and 4-hydroxy-tamoxifen]]		[[Image:Tamoxifen(OH).gif\|right\|thumb\|150px\|chemical structure of tamoxifen and 4-hydroxy-tamoxifen]]

-	~~=== Tamoxifen citrate ===~~	+	One of the uses of [[tamoxifen]] in the lab is the activation of enzymes deactivated via an estrogen-binding domain, like Cre-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains. There are several ways to administer tamoxifen to mice each with advantages and disadvantages. If precise dosage is not paramount, administration via food is the gentlest to the animal. Keep in mind that induction varies depending on Cre and target genes.
-	~~{\| border=1 cellpadding=5 cellspacing=0~~	+
-	\|-	+
-	~~\| molecular weight~~	+
-	~~\| pKa'~~	+
-	~~\| equilibrium solubility in water at 37°C~~	+
-	\|-	+
-	~~\| 563.62 g~~/~~mol~~	+
-	~~\| 8~~.85	+
-	~~\| 0~~.~~5 mg/mL~~	+
-	\|}	+
-	~~see [http://www~~.~~rxlist~~.~~com/cgi/generic/tamox.htm]~~ and ~~[http://pubchem~~.~~ncbi.nlm.nih.gov/summary/summary.cgi?cid=2733525 PubChem entry: tamoxifen citrate]~~	+

-	~~=== 4-hydroxytamoxifen (OHT) ===~~
-	~~Tamoxifen is metabolised to 4-hydroxytamoxifen (often abbreviated OHT).~~
-	~~{\| border=1 cellpadding=5 cellspacing=0~~
-	\|-
-	~~\| molecular weight~~
-	~~\| pKa'~~
-	~~\| equilibrium solubility in water at 37°C~~
-	\|-
-	~~\| 387.51 g/mol~~
-	~~\| ?~~
-	~~\| ? mg/mL~~
-	\|}
-	~~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]~~

	== Tamoxifen administration via chow ==		== Tamoxifen administration via chow ==

Jakob Suckale: categories

2009-04-06T15:06:09Z

Jakob Suckale: links to tamoxifen substrate page

2009-04-06T08:41:40Z

links to tamoxifen substrate page

←Older revision		Revision as of 08:41, 6 April 2009
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	{{back to protocols}}		{{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-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains. There are several ways to administer tamoxifen to mice each with advantages and disadvantages. If precise dosage is not paramount, administration via food is the gentlest to the animal. Keep in mind that induction varies depending on Cre and target genes.	+	One of the uses of [[tamoxifen]] in the lab is the activation of enzymes deactivated via an estrogen-binding domain, like Cre-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains. There are several ways to administer tamoxifen to mice each with advantages and disadvantages. If precise dosage is not paramount, administration via food is the gentlest to the animal. Keep in mind that induction varies depending on Cre and target genes.

-	== Chemical data for tamoxifen ==	+	== Chemical data for [[tamoxifen]] ==

	[[Image:Tamoxifen(OH).gif\|right\|thumb\|150px\|chemical structure of tamoxifen and 4-hydroxy-tamoxifen]]		[[Image:Tamoxifen(OH).gif\|right\|thumb\|150px\|chemical structure of tamoxifen and 4-hydroxy-tamoxifen]]

Jakob Suckale: /* 4-hydroxytamoxifen (OHT) */

2009-04-03T13:17:41Z

4-hydroxytamoxifen (OHT)

←Older revision		Revision as of 13:17, 3 April 2009
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	\| equilibrium solubility in water at 37°C		\| equilibrium solubility in water at 37°C
	\|-		\|-
-	\| ~~563~~.65 g/mol	+	\| 387.51 g/mol
	\| ?		\| ?
	\| ? mg/mL		\| ? mg/mL
	\|}		\|}
	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]
-

	== Tamoxifen administration via chow ==		== Tamoxifen administration via chow ==

Jakob Suckale: intro

2008-09-17T15:44:52Z

intro

←Older revision		Revision as of 15:44, 17 September 2008
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	{{back to protocols}}		{{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-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains.	+	One of the uses of tamoxifen in the lab is the activation of enzymes deactivated via an estrogen-binding domain, like Cre-ER<sup>T</sup> (Cre-LBD). Tamoxifen is an estrogen analogue that binds with higher affinity than estrogen to altered estrogen binding domains. There are several ways to administer tamoxifen to mice each with advantages and disadvantages. If precise dosage is not paramount, administration via food is the gentlest to the animal. Keep in mind that induction varies depending on Cre and target genes.
-		+
-	~~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 data for tamoxifen ==

Jakob Suckale at 15:41, 17 September 2008

2008-09-17T15:41:06Z

←Older revision		Revision as of 15:41, 17 September 2008
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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
		+

	== Tamoxifen administration using a feeding needle ==		== Tamoxifen administration using a feeding needle ==
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	see also Metzger and Chambon, 2001, PMID 11327805, p73		see also Metzger and Chambon, 2001, PMID 11327805, p73
		+

	== Tamoxifen administration by intraperitoneal injection ==		== Tamoxifen administration by intraperitoneal injection ==
	* <font color=green>'''pro''': reliable dosage</font>		* <font color=green>'''pro''': reliable dosage</font>
	* <font color=red>'''con''': probably the most stressful method for the mouse; requires proficient handler</font>		* <font color=red>'''con''': probably the most stressful method for the mouse; requires proficient handler</font>
		+

	== See also ==		== See also ==

Jakob Suckale: pro/con

2008-09-17T15:34:37Z

pro/con

←Older revision		Revision as of 15:34, 17 September 2008
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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]
		+

	== Tamoxifen administration via chow ==		== Tamoxifen administration via chow ==
		+	* <font color=green>'''pro''': less labour-intensive and less stressful for the animals</font>
		+	* <font color=red>'''con''': dosage depends on eating behaviour; induction may take longer then IP</font>
		+
	* 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.
-	* '''advantages''': less labour-intensive and less stressful for the animals

-	== Tamoxifen administration via ~~the~~ drinking water ==	+
		+	== Tamoxifen administration via drinking water ==
		+	* <font color=green>'''pro''': less labour-intensive and less stressful for the animals</font>
		+	* <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>
		+
	#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

-	== Tamoxifen administration ~~via~~ a feeding needle ==	+	== Tamoxifen administration using a feeding needle ==
		+	* <font color=green>'''pro''': dosage more precise than feeding and drinking (less than IP)</font>
		+	* <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.
Line 63:		Line 72:

	== Tamoxifen administration by intraperitoneal injection ==		== Tamoxifen administration by intraperitoneal injection ==
-	* '''con''': probably the most stressful method for the mouse; requires proficient handler	+	* <font color=green>'''pro''': reliable dosage</font>
-	* '''pro''': reliable dosage	+	* <font color=red>'''con''': probably the most stressful method for the mouse; requires proficient handler</font>
-		+

	== See also ==		== See also ==

←Older revision		Revision as of 13:02, 7 April 2009
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	== See also ==		== See also ==
		+	* [[Tamoxifen\|Tamoxifen - chemical information, mechanism of action, ER-deactivated fusion enzymes, stability & storage]]
	* [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]

←Older revision		Revision as of 15:06, 6 April 2009
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	* [http://www.bu.edu/research/compliance/lacu/lacf/sop/injection-techniques/intraperitoneal.shtml Intraperitoneal injection into the mouse] by Boston Uni		* [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		* [http://www.ahwla.org.uk/site/tutorials/BVA/BVA05-Mouse/Mouse.html Mouse restraining and injection] by AHWLA including a video
		+
		+
		+	[[Category:Protocol]]
		+	[[Category:In vivo]]
		+	[[Category:Mouse]]