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Molecular and Cellular Biology, February 2004, p. 1655-1666, Vol. 24, No. 4
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.4.1655-1666.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Mouse Strains with an Active H2-Ea Meiotic Recombination Hot Spot Exhibit Increased Levels of H2-Ea-Specific DNA Breaks in Testicular Germ Cells

Jian Qin,^1, Laura L. Richardson,^2,, Maria Jasin,³ Mary Ann Handel,^2, and Norman Arnheim^1*

Program in Molecular and Computational Biology, University of Southern California, Los Angeles, California 90089-1340,¹ Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee 37996-0840,² Cell Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021³

Received 20 August 2003/ Returned for modification 26 September 2003/ Accepted 19 November 2003

We devised a sensitive method for the site-specific detection of rare meiotic DNA strand breaks in germ cell-enriched testicular cell populations from mice that possess or lack an active recombination hot spot at the H2-Ea gene. Using germ cells from adult animals, we found an excellent correlation between the frequency of DNA breaks in the 418-bp H2-Ea hot spot and crossover activity. The temporal appearance of DNA breaks was also studied in 7- to 18-day-old mice with an active hot spot during the first waves of spermatogenesis. The number of DNA breaks detected rose as leptotene and zygotene spermatocytes populate the testis with a peak at day 14 postpartum, when leptotene, zygotene, and early pachytene spermatocytes are the most common meiotic prophase I cell types. The number of DNA breaks drops precipitously 1 day later, when middle to late pachytene spermatocytes become the dominant subtype. The recombination-related breaks in the hot spot likely reflect SPO11-induced double-strand breaks and/or recombination intermediates containing free 3' hydroxyl groups.

J.Q. and L.L.R. contributed equally to this work.

Present address: Department of Anatomy, Cell and Neurobiology, Joan C. Edwards School of Medicine, Marshall University, Huntington, WV 25704.

Present address: The Jackson Laboratory, Bar Harbor, ME 04609.

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