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First published online 3 May 2005
doi: 10.1242/jcs.02362

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SYCP2 and SYCP3 are required for cohesin core integrity at diplotene but not for centromere cohesion at the first meiotic division

¹ Department of Cell and Molecular Biology and Center for Genomics and Bioinformatics, Karolinska Institutet, Stockholm, 171 77, Sweden
² Department of Gene and Cell Medicine, Mount Sinai School of Medicine, New York, NY 10029, USA

View larger version (75K): [in a new window]   Fig. 1. The absence of SYCP3 results in disassembly of the STAG3 cohesin core. Wild-type (A,B,E,F,I,J) or Sycp3–/– (C,D,G,H,K,L) oocytes were spread and triple-stained with antisera against STAG3 (red), SYCP1 (green) and CREST (white); the stages of meiosis are indicated to the left. An integral axial core labelled by STAG3 antiserum is observed in wild-type pachytene (A,B) and late diplotene (E,F) oocytes. By contrast, the STAG axial core is prematurely fragmented at late diplotene in Sycp3-deficient oocytes (G,H). Yellow labelling indicates co-staining of STAG3 and SYCP1. Arrows in G,H mark STAG3 filaments that are retained in regions that have not yet desynapsed. Bars, 10 µm.

View larger version (27K): [in a new window]   Fig. 2. STAG3 cores and SYCP3 filaments colocalize and are fragmented simultaneously at the early dictyate stage in wild-type oocytes. Oocytes in the early dictyate stage (from newborn animals) were labelled with antisera against SYCP3 (red) and STAG3 (green). A merged picture (centre) shows overlap between the two proteins (yellow). White arrows indicate colocalization of STAG3 and SYCP3, whereas red arrowheads indicate large SYCP3 aggregates, presumably representing self-assembled complexes of dissociated but not yet degraded SYCP3 protein. Bars, 10 µm.

View larger version (87K): [in a new window]   Fig. 3. Fragmentation of the cohesin core occurs at diplotene in the absence of the SYCP3. Late-diplotene oocytes were stained with antisera against SYCP1 (B,D, green; F,H,J,L, red) and CREST (white) in combination with anti-SMC1ß (red), anti-REC8 (green) and anti-SMC3 (green) antibodies. (A,B,E,F,I,J) Wild-type oocytes. (C,D,G,H,K,L) Sycp3–/– oocytes; arrows indicate some of the centromeres. Notice the persistence of cohesin signal at all centromeres (see supplementary material, Fig. S2 for more detail). Bars, 10 µm.

View larger version (76K): [in a new window]   Fig. 4. Cohesin and condensin localization at MI is unaffected in oocytes derived from Sycp3–/– oocytes. The MI oocytes were labelled with antibodies against STAG3 (red) (A-H), REC8 (red) (I,J,M,N), SMC1ß (red) (K,L,O,P), SMC4 (green) (Q-T) or CREST (A-P, green; Q-T, red). (A-D,I-L,Q,R) Wild-type oocytes. (E-H,M-P,S,T) Sycp3–/– oocytes. Bivalents at higher magnification are shown in C-F,I-P. Bars, 5 µm (C-F,I-P), 10 µm (A,B,G,H,Q-T).

View larger version (89K): [in a new window]   Fig. 5. Cohesin distribution and centromere cohesion at MI is unaffected in Sycp3–/– spermatocytes. Cells from wild-type (A,B,J) or Sycp3–/– (C-I,K-M) testes were treated with okadaic acid. Spermatocytes at MI were stained with DAPI (A,C,E,H), or an anti-STAG3 antibody (B,D,M), anti-REC8 antibody (F), anti-SMC1ß antibody (I), anti-SYCP2 antibody (J,K) and anti-CREST (B,D,F,I-K,M). Wild-type MI spermatocytes exhibit 20 bivalents with paired (single) centromeres (B, arrowheads). Sycp3–/– MI spermatocytes have 40 univalents with paired centromeres (D,F,I, arrowheads). SYCP2 fails to accumulate at the centromeres in the absence of SYCP3 (K). Mitotic cells (L,M) derived from Sycp3–/– testes (negative for STAG3) show separated centromere pairs (M, arrows). Bars, 10 µm.

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