A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes
Bivalve haemocytes are essential in defence mechanisms including phagocytosis. They also produce molecules including hydrolytic enzymes and antimicrobial peptides that contribute to pathogen destruction. Although haemocyte activities have been extensively studied, relatively little is known about th...
Published in: | Fish & Shellfish Immunology |
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Online Access: | https://archimer.ifremer.fr/doc/2006/publication-935.pdf https://doi.org/10.1016/j.fsi.2005.06.008 https://archimer.ifremer.fr/doc/00000/935/ |
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ftarchimer:oai:archimer.ifremer.fr:935 2023-05-15T15:57:42+02:00 A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes Aton, Emilie Renault, Tristan Gagnaire, Beatrice Thomas Guyon, Hélène Cognard, C Imbert, N 2006-04 application/pdf https://archimer.ifremer.fr/doc/2006/publication-935.pdf https://doi.org/10.1016/j.fsi.2005.06.008 https://archimer.ifremer.fr/doc/00000/935/ eng eng Elsevier https://archimer.ifremer.fr/doc/2006/publication-935.pdf doi:10.1016/j.fsi.2005.06.008 https://archimer.ifremer.fr/doc/00000/935/ 2005 Elsevier Ltd All rights reserved info:eu-repo/semantics/openAccess restricted use Fish & Shellfish Immunology (1050-4648) (Elsevier), 2006-04 , Vol. 20 , N. 4 , P. 493-502 Cellular activities Fluo 3/AM Calcium Flow cytometry Haemocytes Crassostrea gigas text Publication info:eu-repo/semantics/article 2006 ftarchimer https://doi.org/10.1016/j.fsi.2005.06.008 2021-09-23T20:13:28Z Bivalve haemocytes are essential in defence mechanisms including phagocytosis. They also produce molecules including hydrolytic enzymes and antimicrobial peptides that contribute to pathogen destruction. Although haemocyte activities have been extensively studied, relatively little is known about the intracellular signalling pathways that are evoked during haemocyte activation and especially the role of calcium. Flow cytometry has been used for the first time to define the effect of cell incubation in haemolymph and artificial sea water (ASW) on Pacific oyster, Crassostrea gigas, haemocytes. Cell viability, enzymatic activities (esterases and amino peptidases), phagocytosis and granulocyte percentage were analysed. Viability and some activities were different in haemolymph and ASW. Cytoplasmic-free calcium in circulating haemocytes was then investigated by flow cytometry in both media using a calcium probe (Fluo-3/AM). To explore calcium homeostasis, different calcium modulators were tested. The calcium chelator Bapta/AM (10 mu M) reduced significantly the percentage of Fluo-3-positive cells in ASW. In addition, ryanodine (5 mu M) induced a significant enhancement of the percentage of Fluo-3 positive cells in haemolymph and in ASW. Flow cytometry may be used to study calcium movements in C. gigas haemocytes, but several haemocyte incubation media need to be tested in order to confirm results. The objective of the study should be considered before selecting a particular experimental medium. Article in Journal/Newspaper Crassostrea gigas Pacific oyster Archimer (Archive Institutionnelle de l'Ifremer - Institut français de recherche pour l'exploitation de la mer) Pacific Fish & Shellfish Immunology 20 4 493 502 |
institution |
Open Polar |
collection |
Archimer (Archive Institutionnelle de l'Ifremer - Institut français de recherche pour l'exploitation de la mer) |
op_collection_id |
ftarchimer |
language |
English |
topic |
Cellular activities Fluo 3/AM Calcium Flow cytometry Haemocytes Crassostrea gigas |
spellingShingle |
Cellular activities Fluo 3/AM Calcium Flow cytometry Haemocytes Crassostrea gigas Aton, Emilie Renault, Tristan Gagnaire, Beatrice Thomas Guyon, Hélène Cognard, C Imbert, N A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes |
topic_facet |
Cellular activities Fluo 3/AM Calcium Flow cytometry Haemocytes Crassostrea gigas |
description |
Bivalve haemocytes are essential in defence mechanisms including phagocytosis. They also produce molecules including hydrolytic enzymes and antimicrobial peptides that contribute to pathogen destruction. Although haemocyte activities have been extensively studied, relatively little is known about the intracellular signalling pathways that are evoked during haemocyte activation and especially the role of calcium. Flow cytometry has been used for the first time to define the effect of cell incubation in haemolymph and artificial sea water (ASW) on Pacific oyster, Crassostrea gigas, haemocytes. Cell viability, enzymatic activities (esterases and amino peptidases), phagocytosis and granulocyte percentage were analysed. Viability and some activities were different in haemolymph and ASW. Cytoplasmic-free calcium in circulating haemocytes was then investigated by flow cytometry in both media using a calcium probe (Fluo-3/AM). To explore calcium homeostasis, different calcium modulators were tested. The calcium chelator Bapta/AM (10 mu M) reduced significantly the percentage of Fluo-3-positive cells in ASW. In addition, ryanodine (5 mu M) induced a significant enhancement of the percentage of Fluo-3 positive cells in haemolymph and in ASW. Flow cytometry may be used to study calcium movements in C. gigas haemocytes, but several haemocyte incubation media need to be tested in order to confirm results. The objective of the study should be considered before selecting a particular experimental medium. |
format |
Article in Journal/Newspaper |
author |
Aton, Emilie Renault, Tristan Gagnaire, Beatrice Thomas Guyon, Hélène Cognard, C Imbert, N |
author_facet |
Aton, Emilie Renault, Tristan Gagnaire, Beatrice Thomas Guyon, Hélène Cognard, C Imbert, N |
author_sort |
Aton, Emilie |
title |
A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes |
title_short |
A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes |
title_full |
A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes |
title_fullStr |
A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes |
title_full_unstemmed |
A flow cytometric approach to study intracellular-free Ca2+ in Crassostrea gigas haemocytes |
title_sort |
flow cytometric approach to study intracellular-free ca2+ in crassostrea gigas haemocytes |
publisher |
Elsevier |
publishDate |
2006 |
url |
https://archimer.ifremer.fr/doc/2006/publication-935.pdf https://doi.org/10.1016/j.fsi.2005.06.008 https://archimer.ifremer.fr/doc/00000/935/ |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Crassostrea gigas Pacific oyster |
genre_facet |
Crassostrea gigas Pacific oyster |
op_source |
Fish & Shellfish Immunology (1050-4648) (Elsevier), 2006-04 , Vol. 20 , N. 4 , P. 493-502 |
op_relation |
https://archimer.ifremer.fr/doc/2006/publication-935.pdf doi:10.1016/j.fsi.2005.06.008 https://archimer.ifremer.fr/doc/00000/935/ |
op_rights |
2005 Elsevier Ltd All rights reserved info:eu-repo/semantics/openAccess restricted use |
op_doi |
https://doi.org/10.1016/j.fsi.2005.06.008 |
container_title |
Fish & Shellfish Immunology |
container_volume |
20 |
container_issue |
4 |
container_start_page |
493 |
op_container_end_page |
502 |
_version_ |
1766393391801696256 |