Cnidarian Primary Cell Culture as a Tool to Investigate the Effect of Thermal Stress at Cellular Level
In the context of global change, symbiotic cnidarians are largely affected by seawater temperature elevation leading to symbiosis breakdown. This process, also called bleaching, is triggered by the dysfunction of the symbiont photosystems causing an oxidative stress and cell death to both symbiont and host cells. In our study, we wanted to elucidate the intrinsic capacity of isolated animal cells to deal with thermal stress in the absence of symbiont. In that aim, we have characterized an animal primary cell culture form regenerating tentacles of the temperate sea anemone Anemonia viridis. We first compared the potential of whole tissue tentacle or separated epidermal or gastrodermal monolayers as tissue sources to settle animal cell cultures. Interestingly, only isolated cells extracted from whole tentacles allowed establishing a viable and proliferative primary cell culture throughout 31 days. The analysis of the expression of tissue-specific and pluripotency markers defined cultivated cells as differentiated cells with gastrodermal origin. The characterization of the animal primary cell culture allowed us to submit the obtained gastrodermal cells to hyperthermal stress (+ 5 and + 8 °C) during 1 and 7 days. Though cell viability was not affected at both hyperthermal stress conditions, cell growth drastically decreased. In addition, only a + 8 °C hyperthermia induced a transient increase of antioxidant defences at 1 day but no ubiquitin or carbonylation protein damages. These results demonstrated an intrinsic resistance of cnidarian gastrodermal cells to hyperthermal stress and then confirmed the role of symbionts in the hyperthermia sensitivity leading to bleaching.
KeywordsSea anemones Monolayers Cell differentiation Pluripotency Environmental stress Hyperthermia
Authors greatly acknowledge Laura Hedon for her precious technical help and Thamilla Zamoum for RT-PCR technical advises. Authors also thank Maeva Gesson and Magali Mondin of the PRISM (Platform of Resources in Imaging and Scientific Microscopy, Institut de Biologie Valrose, Université Nice Sophia Antipolis), Matthieu Rouleau and Aldine Amiel for scientific discussions. Authors are also grateful to Brigitte Poderini, for sea anemone maintenance in aquaria. We thank the editor and the reviewers for their useful criticisms, which helped us improve the manuscript.
This work was supported by a doctoral fellowship from the French ministère de l’Enseignement supérieur et de la Recherche (513-EDSFA021-2013) to PV and by research funding program from Université Nice Sophia Antipolis and Provence Alpes Côte d’Azur Region (MIRACLE project).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
- Downs CA, McDougall KE, Woodley CM, Fauth JE, Richmond RH, Kushmaro A, Gibb SW, Loya Y, Ostrander GK, Kramarsky-Winter E (2013) Heat-stress and light-stress induce different cellular pathologies in the symbiotic dinoflagellate during coral bleaching. PLoS One 8(12):e77173CrossRefPubMedPubMedCentralGoogle Scholar
- Furla P, Allemand D, Orsenigo M-N (2000) Involvement of H+-ATPase and carbonic anhydrase in inorganic carbon uptake for endosymbiont photosynthesis. Am J Phys Regul Integr Comp Phys 278:R870–R881Google Scholar
- Higashikubo R, White RA, Roti Roti JL (1993) Flow cytometric BrdUrd-pulse-chase study of heat-induced cell-cycle progression delays. Cell Prolif 26(4):337–348Google Scholar
- Hoegh-Guldberg O, Mumby PJ, Hooten AJ, Steneck RS, Greenfield P, Gomez E, Harvell CD, Sale PF, Edwards AJ, Caldeira K, Knowlton N, Eakin CM, Iglesias-Prieto R, Muthiga N, Bradbury RH, Dubi A, Hatziolos ME (2007) Coral reefs under rapid climate change and ocean acidification. Science 318(5857):1737–1742CrossRefPubMedGoogle Scholar
- Putnam NH, Srivastava M, Hellsten U, Dirks B, Chapman J, Salamov A, Terry A, Shapiro H, Lindquist E, Kapitonov VV, Jurka J, Genikhovich G, Grigoriev IV, Lucas SM, Steele RE, Finnerty JR, Technau U, Martindale MQ, Rokhsar DS (2007) Sea anemone genome reveals ancestral eumetazoan gene repertoire and genomic organization. Science 317(5834):86–94CrossRefPubMedGoogle Scholar
- Richier S, Rodriguez-Lanetty M, Schnitzler CE, Weis VM (2008) Response of the symbiotic cnidarian Anthopleura elegantissima transcriptome to temperature and UV increase. Comp Biochem Physiol Part D Genomics Proteomics 3(4):283–289Google Scholar
- Trench RK (1993) Microalgal-invertebrate symbioses-a review. Endocytobiosis Cell Res 9:135–175Google Scholar
- Vandepas LE, Warren KJ, Amemiya CT, Browne WE (2017) Establishing and maintaining primary cell cultures derived from the ctenophore Mnemiopsis leidyi. J Exp Biol. https://doi.org/10.1242/jeb.152371