Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice
Abstract Transforming acidic acid coiled-coil protein 3 (TACC3) and cytoskeleton associated protein 5 (cKAP5; or colonic hepatic tumor overexpressed gene, chTOG) are vital for spindle assembly and stabilization initiated through TACC3 Aurora-A kinase interaction. Here, TACC3 and cKAP5/chTOG localiza...
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| Format: | Article |
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Nature Portfolio
2024-02-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-55376-z |
| _version_ | 1797274600599453696 |
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| author | Calvin Simerly Emily Robertson Caleb Harrison Sydney Ward Charlize George Jasmine Deleon Carrie Hartnett Gerald Schatten |
| author_facet | Calvin Simerly Emily Robertson Caleb Harrison Sydney Ward Charlize George Jasmine Deleon Carrie Hartnett Gerald Schatten |
| author_sort | Calvin Simerly |
| collection | DOAJ |
| description | Abstract Transforming acidic acid coiled-coil protein 3 (TACC3) and cytoskeleton associated protein 5 (cKAP5; or colonic hepatic tumor overexpressed gene, chTOG) are vital for spindle assembly and stabilization initiated through TACC3 Aurora-A kinase interaction. Here, TACC3 and cKAP5/chTOG localization with monospecific antibodies is investigated in eGFP-centrin-2- expressing mouse meiotic spermatocytes. Both proteins bind spermatocyte spindle poles but neither kinetochore nor interpolar microtubules, unlike in mitotic mouse fibroblasts or female meiotic oocyte spindles. Spermatocytes do not display a liquid-like spindle domain (LISD), although fusing them into maturing oocytes generates LISD-like TACC3 condensates around sperm chromatin but sparse microtubule assembly. Microtubule inhibitors do not reduce TACC3 and cKAP5/chTOG spindle pole binding. MLN 8237 Aurora-A kinase inhibitor removes TACC3, not cKAP5/chTOG, disrupting spindle organization, chromosome alignment, and impacting spindle pole γ-tubulin intensity. The LISD disruptor 1,6-hexanediol abolished TACC3 in spermatocytes, impacting spindle bipolarity and chromosome organization. Cold microtubule disassembly and rescue experiments in the presence of 1,6-hexanediol reinforce the concept that spermatocyte TACC3 spindle pole presence is not required for spindle pole microtubule assembly. Collectively, meiotic spermatocytes without a LISD localize TACC3 and cKAP5/chTOG exclusively at spindle poles to support meiotic spindle pole stabilization during male meiosis, different from either female meiosis or mitosis. |
| first_indexed | 2024-03-07T15:00:41Z |
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| id | doaj.art-ae179d09c22641ce9f1bce1db1bdc8d0 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-07T15:00:41Z |
| publishDate | 2024-02-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj.art-ae179d09c22641ce9f1bce1db1bdc8d02024-03-05T19:09:31ZengNature PortfolioScientific Reports2045-23222024-02-0114111910.1038/s41598-024-55376-zMale meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in miceCalvin Simerly0Emily Robertson1Caleb Harrison2Sydney Ward3Charlize George4Jasmine Deleon5Carrie Hartnett6Gerald Schatten7Departments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterDepartments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterDepartments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterDepartments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterDepartments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterDepartments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterDepartments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterDepartments of Cell Biology, Ob-Gyn-Repro Sci, and Bioengineering, Pittsburgh Development Center of Magee-Womens Research Institute, University of Pittsburgh Medical CenterAbstract Transforming acidic acid coiled-coil protein 3 (TACC3) and cytoskeleton associated protein 5 (cKAP5; or colonic hepatic tumor overexpressed gene, chTOG) are vital for spindle assembly and stabilization initiated through TACC3 Aurora-A kinase interaction. Here, TACC3 and cKAP5/chTOG localization with monospecific antibodies is investigated in eGFP-centrin-2- expressing mouse meiotic spermatocytes. Both proteins bind spermatocyte spindle poles but neither kinetochore nor interpolar microtubules, unlike in mitotic mouse fibroblasts or female meiotic oocyte spindles. Spermatocytes do not display a liquid-like spindle domain (LISD), although fusing them into maturing oocytes generates LISD-like TACC3 condensates around sperm chromatin but sparse microtubule assembly. Microtubule inhibitors do not reduce TACC3 and cKAP5/chTOG spindle pole binding. MLN 8237 Aurora-A kinase inhibitor removes TACC3, not cKAP5/chTOG, disrupting spindle organization, chromosome alignment, and impacting spindle pole γ-tubulin intensity. The LISD disruptor 1,6-hexanediol abolished TACC3 in spermatocytes, impacting spindle bipolarity and chromosome organization. Cold microtubule disassembly and rescue experiments in the presence of 1,6-hexanediol reinforce the concept that spermatocyte TACC3 spindle pole presence is not required for spindle pole microtubule assembly. Collectively, meiotic spermatocytes without a LISD localize TACC3 and cKAP5/chTOG exclusively at spindle poles to support meiotic spindle pole stabilization during male meiosis, different from either female meiosis or mitosis.https://doi.org/10.1038/s41598-024-55376-z |
| spellingShingle | Calvin Simerly Emily Robertson Caleb Harrison Sydney Ward Charlize George Jasmine Deleon Carrie Hartnett Gerald Schatten Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice Scientific Reports |
| title | Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice |
| title_full | Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice |
| title_fullStr | Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice |
| title_full_unstemmed | Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice |
| title_short | Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice |
| title_sort | male meiotic spindle poles are stabilized by tacc3 and ckap5 chtog differently from female meiotic or somatic mitotic spindles in mice |
| url | https://doi.org/10.1038/s41598-024-55376-z |
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