|
|
|
|
 |
|||
|
|||
Phenobarbital-induced hepatocellular proliferation: anti- bromodeoxyuridine and anti-proliferating cell nuclear antigen immunocytochemistry
HB Jones, NA Clarke and NC Barrass
Safety of Medicines Department, ICI Pharmaceuticals, Macclesfield, Cheshire, United Kingdom.
We report modifications to immunocytochemical detection procedures for proliferating cell nuclear antigen (PCNA) which permit its identification in liver samples previously fixed for BrdU immunocytochemistry. Both methods have been used for the assessment of phenobarbital-induced cell proliferation in rat liver. The difficulties associated with the hitherto unsuccessful application of PCNA immunocytochemical methods to tissues fixed in formalin for BrdU visualization were overcome by epitope unmasking with acid hydrolysis, extension of primary antiserum (PC10) incubation, and employment of streptavidin-ABC-HRP. BrdU delivery via osmotic minipumps for 48 hr before euthanasia, followed by fixation in cold formalin for 14 days, yielded reliable and reproducible hepatocellular labeling and a peak of cell proliferation in all lobes on Day 3 (i.e., labeling during Days 1- 3) of dosing with 80 mg/kg/day phenobarbital. Labeling indices (LI) of both control and phenobarbital-treated liver were lower in the left and right median lobes as compared with the lateral lobes. In sections of the left lateral lobe from the same liver, PCNA immunocytochemistry revealed a peak of proliferative activity (about one third of the maximum LI generated by BrdU incorporation) on Day 1. These findings, together with the advantages and disadvantages of both techniques, are discussed in the context of their applications to different investigative requirements.
Volume 41,
Issue 1,
pp. 21-27,
01/01/1993
Copyright © 1993 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  C. Jomary and S. E. Jones Induction of Functional Photoreceptor Phenotype by Exogenous Crx Expression in Mouse Retinal Stem Cells Invest. Ophthalmol. Vis. Sci., January 1, 2008; 49(1): 429 - 437. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. A. Irving, S. S. Shushanov, R. N. Pike, E. Y. Popova, D. Bromme, T. H. T. Coetzer, S. P. Bottomley, I. A. Boulynko, S. A. Grigoryev, and J. C. Whisstock Inhibitory Activity of a Heterochromatin-associated Serpin (MENT) against Papain-like Cysteine Proteinases Affects Chromatin Structure and Blocks Cell Proliferation J. Biol. Chem., April 5, 2002; 277(15): 13192 - 13201. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N.J. Hodges, T.C. Orton, A.J. Strain, and J.K. Chipman Potentiation of epidermal growth factor-induced DNA synthesis in rat hepatocytes by phenobarbitone: possible involvement of oxidative stress and kinase activation Carcinogenesis, November 1, 2000; 21(11): 2041 - 2047. [Abstract] [Full Text] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |