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First published online 12 September 2006
doi: 10.1242/jcs.03182

Journal of Cell Science 119, 4025-4032 (2006)
Published by The Company of Biologists 2006
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Characterization of a novel meiosis-specific protein within the central element of the synaptonemal complex

Geert Hamer1, Katarina Gell1, Anna Kouznetsova1, Ivana Novak1, Ricardo Benavente2 and Christer Höög1,*

1 Department of Cell and Molecular Biology, Karolinska Institute, Berzelius väg 35, Stockholm, SE-171 77, Sweden
2 Department of Cell and Developmental Biology, Biocenter of the University of Würzburg, Am Hubland, Würzburg 97074, Germany


Figure 1
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  		Fig. 1. Identification of TEX12. (A) In situ hybridization using Sycp3 and Tex12 on sections of mouse testes (A) or ovaries (B). (C) Western blot analysis of TEX12 using a somatic cell line (C2C12) and mouse testis lysates. (D) Immunohistochemical localization of TEX12 in the testis; arrows indicate examples of histochemically (brown) stained cells. pL, pre-leptotene spermatocytes; Ls, leptotene spermatocytes; Ps, pachytene spermatocytes; Rs, round spermatocytes; Ser, Sertoli cell; Ley, Leydig cells. Stages of the seminiferous epithelium are indicated with roman numerals. Bars, 100 µm (A); 20 µm (B,D).

 

Figure 2
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  		Fig. 2. TEX12 specifically localizes to the synaptonemal complex of synapsed homologous chromosomes. (A) Spermatocytes stained for SYCP3 (green), TEX12 (red) and CREST (white in the merged images). (B) Spermatocytes stained for SYCP1 (green), TEX12 (red) and CREST (white in the merged images). Bars, 5 µm.

 

Figure 3
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  		Fig. 3. TEX12 specifically localizes to the central element structure of the synaptonemal complex. Electron microscopy showing immunogold labelling of TEX12 (A) and the coiled-coil region of SYCP1 (B). Bars, 0.1 µm.

 

Figure 4
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  		Fig. 4. TEX12 co-localizes with SYCE2, but not necessarily with SYCE1 or SYCP1. Pachytene spermatocytes stained for SYCE1 or SYCE2 (green), TEX12 (red) and CREST (white in the merged images). Bar, 5 µm.

 

Figure 5
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  		Fig. 5. SYCP1 is required for TEX12 localization to the synapsed axes. (A) Sycp3-/- spermatocytes stained for TEX12 (red), the meiotic cohesin STAG3 (green) and CREST (white in the merged image). (B) Sycp1-/- spermatocytes stained for TEX12, SYCE1 or SYCE2 (red), SYCP3 (green) and CREST (white in the merged images). Bars, 5 µm.

 

Figure 6
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  		Fig. 6. Interaction between TEX12 and SYCE2. Western blot analysis of co-immunoprecipitations (IP) using anti-SYCE1, anti-SYCE2, anti-SYCP1 or rabbit-serum-coupled beads, confirming that SYCE1 co-immunoprecipitates with SYCP1 (Costa et al., 2005Go) and showing that TEX12 co-immunoprecipitates with SYCE2; plus the `reverse' IPs, using TEX12 as bait, showing that SYCE2 co-immunoprecipitates with TEX12.

 




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