title: Lectin-Based Profiling of Coelomocytes in Holothuria scabra and Expression of Superoxide Dismutase in Purified Coelomocytes.
abstract: Coelomocytes are the first line of immune defense in marine animals. Their distributions are greatly variable even in the close animal species. In this study, we used lectin staining to aid in the classification and purification of these cells for further investigation of SOD distribution among coelomocytes of H. scraba. We classified coelomocytes into four types: type 1, lymphocytes; type 2, phagocytes; type 3, spherulocytes; and type 4, giant cells. Among four lectins used, Con A appeared to give a broad reactivity against most coelomocytes, except for giant cells. In addition, phagocytes usually engaged the highest fluorescent intensity with most lectins, with the exception of PNA, for which spherulocytes possessed the highest fluorescent intensity. Using FACS for fraction collection, it was found that F1 fraction contained the purest phagocyte population (> 95%), which was highly reactive with anti- superoxide dismutase (SOD) as revealed by immunoblotting and immunofluorescence staining, although some minor staining was also detected in spherulocytes. Our results thus provide a fundamental platform for comparing alterations that may happen to the population and SOD contents of coelomocytes when the sea cucumber is subjected to environmental changes that would activate their immune responses.
authors.affiliation: 1 Department of Anatomy;
authors.forename: Yanvit
authors.initials: Y
authors.lastname: Prompoon
authors.affiliation: 1 Department of Anatomy;
authors.forename: Wattana
authors.initials: W
authors.lastname: Weerachatyanukul
authors.affiliation: 1 Department of Anatomy;|2 Center of Excellence for Shrimp Molecular Biology and Biotechnology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.|3 Shrimp Genetic Improvement Center, Chaiya, Nakhon Sri Thammarat, 84110, Thailand.
authors.forename: Boonsirm
authors.initials: B
authors.lastname: Withyachumnarnkul
authors.affiliation: 1 Department of Anatomy;|2 Center of Excellence for Shrimp Molecular Biology and Biotechnology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
authors.forename: Rapeepun
authors.initials: R
authors.lastname: Vanichviriyakit
authors.affiliation: 1 Department of Anatomy;
authors.forename: Kanokpan
authors.initials: K
authors.lastname: Wongprasert
authors.affiliation: 1 Department of Anatomy;
authors.forename: Somluk
authors.initials: S
authors.lastname: Asuvapongpatana
mesh_terms: D000818:Animals; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D047332:Holothuria / Q000502:physiology*; D037102:Lectins / Q000502:physiology*; D010586:Phagocytes / Q000166:cytology; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D037102:Lectins; D013482:Superoxide Dismutase
keywords: H. scabra; coelomocytes; immune response; sea cucumber; superoxide dismutase