Re: Circulation time of BRDU in a rat

<< Previous Message | Next Message >> (Jamie Erickson),
Date:Thu, 24 Jun 1999 09:46:43 -0400 (Eastern Daylight Time)


We use the following:

Dissolve 20mg BrdU per 1 ml phosphate buffered  sterile saline (pH 7.6), held at room temp. and
shielded from light.  This is administered IP at 5 ml/1Kg.  The animals are sac.'ed 1 hour after
injection.   So much for the easy part.

As regards measuring proliferation labeling index (LI).  There are several ways that you could
generate the results, number of labeled cells/ mm (or other length unit) or number pos. cells
divided by total cells counted per crypt. we have used both of these methods (still do). see :

The Effects of Exposure Route on DNA Adduct Formation and Cellular Proliferation by
D.La, R. Schoonhoven, N. Ito, J.A. Swenberg; 
Toxicology and Applied Pharmacology 140, 108-114 (1996)

Most of the details (on how we measured the LI) are given in that article.   Our IHC methods have
changed since then (darn that thar  progress thing ;-))  By measuring migration I would set up a
time study over several days, no more than 7 as at that point ALL cells would be labeled. Do an IP
injection each day ie: day 1 sac day 1 , day 2 (rec'd 2 IP injections) sac,  etc...

Measure either length of labeled cells along basement membrane from base of crypt on up or, num. of
cells per measurement unit.

Please feel free to call if I can be of any assistance.  

-- Begin original message --
> HI All,
>       I know this was discussed sometime ago but could someone tell me the circulation time for 
> BRDU, I believe the dose of BRDU given was 500 ug/kg.  I'm doing a study to evaluate BRDU stained 
> slides of Rat colon and need this info. Also how do people evaluate these slides, is it by number 
> of positive cells/ crypt (how many do you count /slides?) . There seems to be two things to 
> evaluate in these slides migration up the Crypt and proliferation, what's the best way to  
> evaluate both of these??
> If someone could help I'd really appreciate it.
> Jamie Erickson

-- End original message --

best regards,
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
Dept. of Environmental Sciences and Engineering
University of North Carolina
Chapel Hill, NC 27599
office 919-966-6343
   Lab 919-966-6140
   Fax 919-966-6123 

Don't go around saying the world owes you a living; the world owes you
nothing; it was here first. 
Mark Twain [Samuel Langhornne Clemens] (1835-1910) 

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