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. Author manuscript; available in PMC: 2016 Jan 21.
Published in final edited form as: Science. 2002 Apr 4;296(5569):922–927. doi: 10.1126/science.1069398

Fig. 4. Defective spermatogenesis in H2AX−/− mice.

Fig. 4

(A) Comparison of testis size in 2-month-old H2AX wild-type (+/+) and knockout (−/−) mice. Bar, 2 mm. (B) Sections of seminiferous tubules of 7-week-old (+/+ and −/−) littermates stained with hematoxylin-eosin. Magnification, ×10. (C) Hematoxylin-eosin-stained sections of epididymis from the same mice shown in (B). Magnification, ×40. (D) High-magnification (×100) images of periodic acid–Schiff–stained paraffin sections of seminiferous tubules of 2-month old wild-type (+/+) (left panel) and knockout (−/−) (two right panels) mice. Primary spermatocytes in early pachytene (EP), late pachytene (LP), and zygotype (Z) are indicated. Apoptotic nuclei with condensing chromatin are present in H2AX−/− tubules (arrows). (E) TUNEL (terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling) assay detects very few apoptotic cells in normal tubules (arrows, left panel), whereas H2AX-mutant tubules contain a large number of dying cells. Magnification, ×40. (F to I) Indirect immunofluorescence of H2AX+/− (+/−; left panels) and H2AX−/− (−/−; right panels) pachytene spermatocytes. (F) Merged image of Scp3 (red) and γ-H2AX (green). (G) Merged image of Scp3 (green) and Scp1 (red). (H) Merged image of Scp1 (red) and Mlh1 (green). (I) Diplotene (left) and diakinesis (right) H2AX+/− stages (two individual cells separated by the dotted line), and fragmented synaptonemal complex in H2AX−/− spermatocyte visualized with antibody to Scp3 (red) and counterstained with DAPI (blue). For (F) to (H), the arrowhead indicates the Y chromosome, and the arrow shows the X chromosome. Bar [(F) to (I)], 10 μm.