Examination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopy
Abstract Background Eimeria parasite infection occurs via ingestion of oocysts. The robust, bilayer oocyst wall is formed from the contents of wall-forming bodies (WFBs), WFB1 and WFB2, located exclusively in macrogametocytes. Eimeria necatrix gametocyte proteins 22 and 59 (EnGAM22 and EnGAM59) have...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-04-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13071-023-05742-z |
| _version_ | 1797846090573152256 |
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| author | Lele Wang Dandan Liu Yang Gao Zhaofeng Hou Yu Zhu Feiyan Wang Wenjing Li Amin Zhang Jinjun Xu Junjie Hu Jianping Tao |
| author_facet | Lele Wang Dandan Liu Yang Gao Zhaofeng Hou Yu Zhu Feiyan Wang Wenjing Li Amin Zhang Jinjun Xu Junjie Hu Jianping Tao |
| author_sort | Lele Wang |
| collection | DOAJ |
| description | Abstract Background Eimeria parasite infection occurs via ingestion of oocysts. The robust, bilayer oocyst wall is formed from the contents of wall-forming bodies (WFBs), WFB1 and WFB2, located exclusively in macrogametocytes. Eimeria necatrix gametocyte proteins 22 and 59 (EnGAM22 and EnGAM59) have been found to localize to WFBs and the oocyst wall. However, the exact localization of these two proteins is not clear. Methods WFBs of E. necatrix were extracted from purified gametocytes using a cutoff filter and the extracts of purified WFBs and gametocytes were analyzed using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Then, the localization of EnGAM22 and EnGAM59 proteins was determined using an indirect immunofluorescence assay. Finally, the development of macrogametocytes and the oocyst wall of E. necatrix was analyzed using laser confocal microscopy and scanning electron microscopy. Results Purified WFBs had the same shape and size as those observed in macrogametocytes. EnGAM22 protein localized to WFB1, whereas EnGAM59 protein localized to WFB2. Both EnGAM22 and EnGAM59 native proteins were detected in the extracts of WFBs and gametocytes. The outer layer of the oocyst wall was formed by the release of the contents of WFB1 at the surface of the macrogametocyte to form an anti-EnGAM22 positive layer. WFB2 then appeared to give rise to the inner layer, which was anti-EnGAM59 positive. Conclusions EnGAM22 and EnGAM59 proteins localized to WFB1 and WFB2 and were involved in the formation of the outer and inner layers of the oocyst wall of E. necatrix, respectively. The processes of macrogametogenesis and oocyst wall formation of E. necatrix are similar to other Eimeria parasites. The anti-EnGAM22 antibody could be used as a tool to track the transport and secretion of proteins in WFB1 during oocyst development. Graphical Abstract |
| first_indexed | 2024-04-09T17:49:19Z |
| format | Article |
| id | doaj.art-76c3bff5b088479c995cb2f0df7eb6e9 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-04-09T17:49:19Z |
| publishDate | 2023-04-01 |
| publisher | BMC |
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| series | Parasites & Vectors |
| spelling | doaj.art-76c3bff5b088479c995cb2f0df7eb6e92023-04-16T11:08:04ZengBMCParasites & Vectors1756-33052023-04-0116111210.1186/s13071-023-05742-zExamination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopyLele Wang0Dandan Liu1Yang Gao2Zhaofeng Hou3Yu Zhu4Feiyan Wang5Wenjing Li6Amin Zhang7Jinjun Xu8Junjie Hu9Jianping Tao10College of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversityCollege of Veterinary Medicine, Yangzhou UniversitySchool of Ecology and Environmental Sciences and Yunnan Key Laboratory for Plateau Mountain Ecology and Restoration of Degraded Environments, Yunnan UniversityCollege of Veterinary Medicine, Yangzhou UniversityAbstract Background Eimeria parasite infection occurs via ingestion of oocysts. The robust, bilayer oocyst wall is formed from the contents of wall-forming bodies (WFBs), WFB1 and WFB2, located exclusively in macrogametocytes. Eimeria necatrix gametocyte proteins 22 and 59 (EnGAM22 and EnGAM59) have been found to localize to WFBs and the oocyst wall. However, the exact localization of these two proteins is not clear. Methods WFBs of E. necatrix were extracted from purified gametocytes using a cutoff filter and the extracts of purified WFBs and gametocytes were analyzed using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Then, the localization of EnGAM22 and EnGAM59 proteins was determined using an indirect immunofluorescence assay. Finally, the development of macrogametocytes and the oocyst wall of E. necatrix was analyzed using laser confocal microscopy and scanning electron microscopy. Results Purified WFBs had the same shape and size as those observed in macrogametocytes. EnGAM22 protein localized to WFB1, whereas EnGAM59 protein localized to WFB2. Both EnGAM22 and EnGAM59 native proteins were detected in the extracts of WFBs and gametocytes. The outer layer of the oocyst wall was formed by the release of the contents of WFB1 at the surface of the macrogametocyte to form an anti-EnGAM22 positive layer. WFB2 then appeared to give rise to the inner layer, which was anti-EnGAM59 positive. Conclusions EnGAM22 and EnGAM59 proteins localized to WFB1 and WFB2 and were involved in the formation of the outer and inner layers of the oocyst wall of E. necatrix, respectively. The processes of macrogametogenesis and oocyst wall formation of E. necatrix are similar to other Eimeria parasites. The anti-EnGAM22 antibody could be used as a tool to track the transport and secretion of proteins in WFB1 during oocyst development. Graphical Abstracthttps://doi.org/10.1186/s13071-023-05742-zEimeria necatrixGametocyte proteinsOocyst wall formationWall-forming bodies |
| spellingShingle | Lele Wang Dandan Liu Yang Gao Zhaofeng Hou Yu Zhu Feiyan Wang Wenjing Li Amin Zhang Jinjun Xu Junjie Hu Jianping Tao Examination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopy Parasites & Vectors Eimeria necatrix Gametocyte proteins Oocyst wall formation Wall-forming bodies |
| title | Examination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopy |
| title_full | Examination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopy |
| title_fullStr | Examination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopy |
| title_full_unstemmed | Examination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopy |
| title_short | Examination of gametocyte protein 22 localization and oocyst wall formation in Eimeria necatrix using laser confocal microscopy and scanning electron microscopy |
| title_sort | examination of gametocyte protein 22 localization and oocyst wall formation in eimeria necatrix using laser confocal microscopy and scanning electron microscopy |
| topic | Eimeria necatrix Gametocyte proteins Oocyst wall formation Wall-forming bodies |
| url | https://doi.org/10.1186/s13071-023-05742-z |
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