Apoptosis, FACs and aberrant metaphase II and spermatid cells in Psma8-deficient mice.
(A) Double immunolabeling of Caspase3 (green) and (B) TUNEL (green) with SYCP3 (red). Non-apoptotic metaphase I cells from Psma8+/+ show absence of green staining whereas apoptotic metaphases I from Psma8-/- show intense Caspase-3 and TUNEL labeling. Chromatin was counterstained with DAPI. (C) Acrosome positive labeling of round spermatids by PNA staining (green). (D) FACs analysis of cells from whole seminiferous tubules from wild type and Psma8-/- showing in both genotypes (N = 2) the presence of 4C, 2C and 1C compartment as a result of the early spermatid arrest. Source data describing the gating strategy is shown in S5B Fig. (E) Double immunolabeling of metaphase I cells with tubulin (green) and ACA (red) showing normal (Psma8+/+) and abnormal spindles (Psma8-/-). (F) Double immunolabeling of SYCP3 (green) with ACA (red) in wild-type and Psma8-/- spermatocytes at metaphase II which shows aberrant accumulation of SYCP3 at the centromeres. (G) Double immunolabeling of PA200 (green) and SYCP3 (red) in chromosome spreads. PA200 is detected at the chromosome axes of the autosomal and XY bivalents during pachytene in wild type spermatocytes in contrast to the absence of labeling in Psma8-/- spermatocytes. Bar in panels (C, E) 5 μm and 10 μm (A, B, F and G).