Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin
The vitelline layer (VL) of a sea urchin egg is an intricate meshwork of glycoproteins that intimately ensheathes the plasma membrane. During fertilization, the VL plays important roles. Firstly, the receptors for sperm reside on the VL. Secondly, following cortical granule exocytosis, the VL is ele...
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MDPI AG
2021-12-01
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| Online Access: | https://www.mdpi.com/2073-4409/10/12/3573 |
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| author | Nunzia Limatola Jong Tai Chun Sawsen Cherraben Jean-Louis Schmitt Jean-Marie Lehn Luigia Santella |
| author_facet | Nunzia Limatola Jong Tai Chun Sawsen Cherraben Jean-Louis Schmitt Jean-Marie Lehn Luigia Santella |
| author_sort | Nunzia Limatola |
| collection | DOAJ |
| description | The vitelline layer (VL) of a sea urchin egg is an intricate meshwork of glycoproteins that intimately ensheathes the plasma membrane. During fertilization, the VL plays important roles. Firstly, the receptors for sperm reside on the VL. Secondly, following cortical granule exocytosis, the VL is elevated and transformed into the fertilization envelope (FE), owing to the assembly and crosslinking of the extruded materials. As these two crucial stages involve the VL, its alteration was expected to affect the fertilization process. In the present study, we addressed this question by mildly treating the eggs with a reducing agent, dithiothreitol (DTT). A brief pretreatment with DTT resulted in partial disruption of the VL, as judged by electron microscopy and by a novel fluorescent polyamine probe that selectively labelled the VL. The DTT-pretreated eggs did not elevate the FE but were mostly monospermic at fertilization. These eggs also manifested certain anomalies at fertilization: (i) compromised Ca<sup>2+</sup> signaling, (ii) blocked translocation of cortical actin filaments, and (iii) impaired cleavage. Some of these phenotypic changes were reversed by restoring the DTT-exposed eggs in normal seawater prior to fertilization. Our findings suggest that the FE is not the decisive factor preventing polyspermy and that the integrity of the VL is nonetheless crucial to the egg’s fertilization response. |
| first_indexed | 2024-03-10T04:24:34Z |
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| id | doaj.art-24faba7db4dd49a3bb94b4418968ddd8 |
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| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T04:24:34Z |
| publishDate | 2021-12-01 |
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| spelling | doaj.art-24faba7db4dd49a3bb94b4418968ddd82023-11-23T07:39:41ZengMDPI AGCells2073-44092021-12-011012357310.3390/cells10123573Effects of Dithiothreitol on Fertilization and Early Development in Sea UrchinNunzia Limatola0Jong Tai Chun1Sawsen Cherraben2Jean-Louis Schmitt3Jean-Marie Lehn4Luigia Santella5Department of Research Infrastructures for Marine Biological Resources, Stazione Zoologica Anton Dohrn, 80121 Napoli, ItalyDepartment of Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, 80121 Napoli, ItalyLaboratory of Supramolecular Chemistry, Institut de Science et d’Ingénierie Supramoléculaires ISIS—Université de Strasbourg, 8 allée Gaspard Monge, 67000 Strasbourg, FranceLaboratory of Supramolecular Chemistry, Institut de Science et d’Ingénierie Supramoléculaires ISIS—Université de Strasbourg, 8 allée Gaspard Monge, 67000 Strasbourg, FranceLaboratory of Supramolecular Chemistry, Institut de Science et d’Ingénierie Supramoléculaires ISIS—Université de Strasbourg, 8 allée Gaspard Monge, 67000 Strasbourg, FranceDepartment of Research Infrastructures for Marine Biological Resources, Stazione Zoologica Anton Dohrn, 80121 Napoli, ItalyThe vitelline layer (VL) of a sea urchin egg is an intricate meshwork of glycoproteins that intimately ensheathes the plasma membrane. During fertilization, the VL plays important roles. Firstly, the receptors for sperm reside on the VL. Secondly, following cortical granule exocytosis, the VL is elevated and transformed into the fertilization envelope (FE), owing to the assembly and crosslinking of the extruded materials. As these two crucial stages involve the VL, its alteration was expected to affect the fertilization process. In the present study, we addressed this question by mildly treating the eggs with a reducing agent, dithiothreitol (DTT). A brief pretreatment with DTT resulted in partial disruption of the VL, as judged by electron microscopy and by a novel fluorescent polyamine probe that selectively labelled the VL. The DTT-pretreated eggs did not elevate the FE but were mostly monospermic at fertilization. These eggs also manifested certain anomalies at fertilization: (i) compromised Ca<sup>2+</sup> signaling, (ii) blocked translocation of cortical actin filaments, and (iii) impaired cleavage. Some of these phenotypic changes were reversed by restoring the DTT-exposed eggs in normal seawater prior to fertilization. Our findings suggest that the FE is not the decisive factor preventing polyspermy and that the integrity of the VL is nonetheless crucial to the egg’s fertilization response.https://www.mdpi.com/2073-4409/10/12/3573vitelline layerfertilizationsea urchin eggsplasticityCa<sup>2+</sup> signalingactin |
| spellingShingle | Nunzia Limatola Jong Tai Chun Sawsen Cherraben Jean-Louis Schmitt Jean-Marie Lehn Luigia Santella Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin Cells vitelline layer fertilization sea urchin eggs plasticity Ca<sup>2+</sup> signaling actin |
| title | Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin |
| title_full | Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin |
| title_fullStr | Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin |
| title_full_unstemmed | Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin |
| title_short | Effects of Dithiothreitol on Fertilization and Early Development in Sea Urchin |
| title_sort | effects of dithiothreitol on fertilization and early development in sea urchin |
| topic | vitelline layer fertilization sea urchin eggs plasticity Ca<sup>2+</sup> signaling actin |
| url | https://www.mdpi.com/2073-4409/10/12/3573 |
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