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Volume 51 (12): 1681-1688, 2003
Copyright ©The Histochemical Society, Inc.
Evaluation of Cell Proliferation in Rat Tissues with BrdU, PCNA, Ki-67(MIB-5) Immunohistochemistry and In Situ Hybridization for Histone mRNA
Charles River Laboratories (LM,JRL,EBH) and Division of Biometry and Risk Assessment of National Center for Toxicological Research (RLK), Jefferson, Arkansas
Correspondence to: Levan Muskhelishvili, PhD, Charles River Laboratories at National Center for Toxicological Research, 3900 NCTR Rd., MC 923, Jefferson, AR 72079. E-mail: lmuskhelishvili{at}nctr.fda.gov
The standard method for assessment of cell proliferation in paraffin-embedded tissue sections is 5-bromodeoxyuridine (BrdU) immunohistochemistry (IHC). BrdU can be administered to laboratory animals via IP injections, is readily incorporated into nuclei during the DNA synthetic phase of the cell cycle, and is detected with an anti-BrdU antibody. This method has several disadvantages, and an accurate method for evaluation of proliferative activity that can substitute for BrdU IHC, when necessary, is of great interest to investigators. Alternative methods for detection of proliferating cells in tissue sections are proliferating cell nuclear antigen (PCNA) IHC, Ki-67 IHC, and in situ hybridization (ISH) for histone mRNA. To determine the optimal choice, we analyzed the correlation of anti-PCNA, anti-Ki-67(MIB-5), and histone mRNA labeling indices (LIs) with anti-BrdU LI in rat highly replicative (renewing) tissues. The correlation between anti-BrdU and histone mRNA LIs, as well as the correlation between anti-BrdU and anti-Ki-67 LIs, was statistically significant. There was no significant correlation between anti-BrdU and anti-PCNA LIs. These results suggest that both ISH for histone mRNA and IHC with MIB-5 are preferable techniques for assessment of cell proliferation in rat paraffin-embedded renewing tissues compared to PCNA IHC. They can substitute for BrdU IHC when necessary.
(J Histochem Cytochem 51:1681–1688, 2003)
Key Words: proliferation • immunohistochemistry • in situ hybridization • labeling indices
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