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Scleractinian coral cell proliferation is reduced in primary culture of suspended multicellular aggregates compared to polyps
Authors:A. Lecointe  S. Cohen  M. Gèze  C. Djediat  A. Meibom  I. Domart-Coulon
Affiliation:1. UMR7245 MCAM Département RDDM, Muséum National D’Histoire Naturelle, Case Postale 54, 57 rue Cuvier, 75005, Paris, France
2. Laboratory for Biological Geochemistry, School of Architecture, Civil and Environmental Engineering (ENAC), Ecole Polytechnique Fédérale de Lausanne (EPFL), 1015, Lausanne, Switzerland
3. The Interuniversity Institute for Marine Science in Eilat, P.O. Box 469, 88103, Eilat, Israel
4. The Mina and Everard Goodman Faculty for Life Sciences, Bar-Ilan University, 52900, Ramat Gan, Israel
5. Plateforme de microscopie électronique et d’imagerie, Département RDDM, Muséum National D’Histoire Naturelle, Case Postale 39, 12 rue Buffon, 75005, Paris, France
Abstract:Cell cultures from reef-building scleractinian corals are being developed to study the response of these ecologically important organisms to environmental stress and diseases. Despite the importance of cell division to support propagation, cell proliferation in polyps and in vitro is under-investigated. In this study, suspended multicellular aggregates (tissue balls) were obtained after collagenase dissociation of Pocillopora damicornis coral, with varying yields between enzyme types and brands. Ultrastructure and cell type distribution were characterized in the tissue balls (TBs) compared to the polyp. Morphological evidence of cellular metabolic activity in their ciliated cortex and autophagy in their central mass suggests involvement of active tissue reorganization processes. DNA synthesis was evaluated in the forming multicellular aggregates and in the four cell layers of the polyp, using BrdU labeling of nuclei over a 24 h period. The distribution of BrdU-labeled coral cells was spatially heterogeneous and their proportion was very low in tissue balls (0.2 ± 0.1 %), indicating that suspended multicellular aggregate formation does not involve significant cell division. In polyps, DNA synthesis was significantly lower in the calicoderm (<1 %) compared to both oral and aboral gastroderm (about 10 %) and to the pseudostratified oral epithelium (15–25 % at tip of tentacle). DNA synthesis in the endosymbiotic dinoflagellates dropped in the forming tissue balls (2.7 ± 1.2 %) compared to the polyp (14 ± 3.4 %) where it was not different from the host gastroderm (10.3 ± 1.2 %). A transient (24 h) increase was observed in the cell-specific density of dinoflagellates in individually dissociated coral cell cultures. These results suggest disruption of coral cell proliferation processes upon establishment in primary culture.
Keywords:Primary culture   DNA synthesis   BrdU   Coral   Scleractinia   Dinoflagellate   Cell proliferation
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