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Flow cytometric analysis of micronuclei in peripheral blood reticulocytes III. An efficient method of monitoring chromosomal damage in the beagle dog.
Toxicol Sci. 2007 Dec; 100(2):406-14.TS

Abstract

Erythrocyte-based micronucleus tests have traditionally analyzed bone marrow because splenic filtration in most species removes micronucleated cells from peripheral blood. We have evaluated a flow cytometric method for monitoring micronucleated reticulocyte frequencies (%MN-RET) in the peripheral blood of beagle dogs treated with cyclophosphamide (CP) and have found that analysis of micronucleated reticulocytes (MN-RETs) in peripheral blood is a suitable surrogate for bone marrow analysis. The three-color flow cytometric method uses anti-CD71 labeling to identify reticulocytes and Plasmodium berghei-containing erythrocytes as a calibration standard. The spontaneous %MN-RET determined by flow cytometry was 0.31 +/- 0.09% (n = 22) for peripheral blood, compared with 0.38 +/- 0.13% (SD, n = 12) for bone marrow, and 0.27 +/- 0.08% (n = 12) for peripheral blood by microscopic scoring with acridine orange staining. The kinetics of appearance and disappearance of MN-RETs in blood were determined by collecting daily samples after iv treatment with CP. The maximum frequency occurred approximately 48 h after dosing. Frequencies of MN-RETs in peripheral blood at steady state following daily CP treatment were 55-68% of corresponding bone marrow values assessed by microscopy and 55-112% as assessed by flow cytometry. This difference is presumably due to splenic removal, which appears slightly less stringent than that previously reported for CP-treated Sprague-Dawley rats. Responses in bone marrow and peripheral blood were highly correlated and similar to or greater than those reported in mice and rats at equitoxic doses.

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Authors+Show Affiliations

Harper SB
Department of Veterans Affairs, Washington, DC 20422, USA.
Dertinger SD
No affiliation info available
Bishop ME
No affiliation info available
Lynch AM
No affiliation info available
Lorenzo M
No affiliation info available
Saylor M
No affiliation info available
MacGregor JT
No affiliation info available

MeSH

AnimalsBone MarrowBone Marrow CellsCyclophosphamideDogsFemaleFlow CytometryMaleMicronuclei, Chromosome-DefectiveMicronucleus TestsMutagensReproducibility of ResultsReticulocytes

Pub Type(s)

Comparative Study
Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

17872896

Citation

Harper, Susan B., et al. "Flow Cytometric Analysis of Micronuclei in Peripheral Blood Reticulocytes III. an Efficient Method of Monitoring Chromosomal Damage in the Beagle Dog." Toxicological Sciences : an Official Journal of the Society of Toxicology, vol. 100, no. 2, 2007, pp. 406-14.
Harper SB, Dertinger SD, Bishop ME, et al. Flow cytometric analysis of micronuclei in peripheral blood reticulocytes III. An efficient method of monitoring chromosomal damage in the beagle dog. Toxicol Sci. 2007;100(2):406-14.
Harper, S. B., Dertinger, S. D., Bishop, M. E., Lynch, A. M., Lorenzo, M., Saylor, M., & MacGregor, J. T. (2007). Flow cytometric analysis of micronuclei in peripheral blood reticulocytes III. An efficient method of monitoring chromosomal damage in the beagle dog. Toxicological Sciences : an Official Journal of the Society of Toxicology, 100(2), 406-14.
Harper SB, et al. Flow Cytometric Analysis of Micronuclei in Peripheral Blood Reticulocytes III. an Efficient Method of Monitoring Chromosomal Damage in the Beagle Dog. Toxicol Sci. 2007;100(2):406-14. PubMed PMID: 17872896.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Flow cytometric analysis of micronuclei in peripheral blood reticulocytes III. An efficient method of monitoring chromosomal damage in the beagle dog. AU - Harper,Susan B, AU - Dertinger,Stephen D, AU - Bishop,Michelle E, AU - Lynch,Anthony M, AU - Lorenzo,Maria, AU - Saylor,Michelle, AU - MacGregor,James T, Y1 - 2007/09/13/ PY - 2007/9/18/pubmed PY - 2007/12/21/medline PY - 2007/9/18/entrez SP - 406 EP - 14 JF - Toxicological sciences : an official journal of the Society of Toxicology JO - Toxicol Sci VL - 100 IS - 2 N2 - Erythrocyte-based micronucleus tests have traditionally analyzed bone marrow because splenic filtration in most species removes micronucleated cells from peripheral blood. We have evaluated a flow cytometric method for monitoring micronucleated reticulocyte frequencies (%MN-RET) in the peripheral blood of beagle dogs treated with cyclophosphamide (CP) and have found that analysis of micronucleated reticulocytes (MN-RETs) in peripheral blood is a suitable surrogate for bone marrow analysis. The three-color flow cytometric method uses anti-CD71 labeling to identify reticulocytes and Plasmodium berghei-containing erythrocytes as a calibration standard. The spontaneous %MN-RET determined by flow cytometry was 0.31 +/- 0.09% (n = 22) for peripheral blood, compared with 0.38 +/- 0.13% (SD, n = 12) for bone marrow, and 0.27 +/- 0.08% (n = 12) for peripheral blood by microscopic scoring with acridine orange staining. The kinetics of appearance and disappearance of MN-RETs in blood were determined by collecting daily samples after iv treatment with CP. The maximum frequency occurred approximately 48 h after dosing. Frequencies of MN-RETs in peripheral blood at steady state following daily CP treatment were 55-68% of corresponding bone marrow values assessed by microscopy and 55-112% as assessed by flow cytometry. This difference is presumably due to splenic removal, which appears slightly less stringent than that previously reported for CP-treated Sprague-Dawley rats. Responses in bone marrow and peripheral blood were highly correlated and similar to or greater than those reported in mice and rats at equitoxic doses. SN - 1096-6080 UR - https://www.unboundmedicine.com/medline/citation/17872896/Flow_cytometric_analysis_of_micronuclei_in_peripheral_blood_reticulocytes_III__An_efficient_method_of_monitoring_chromosomal_damage_in_the_beagle_dog_ L2 - https://academic.oup.com/toxsci/article-lookup/doi/10.1093/toxsci/kfm241 DB - PRIME DP - Unbound Medicine ER -