Hostname: page-component-78c5997874-lj6df Total loading time: 0 Render date: 2024-11-01T17:25:46.443Z Has data issue: false hasContentIssue false
  • English
  • Français

Osmolarity and composition of cell culture media affect further development and survival in zebrafish embryos

Published online by Cambridge University Press:  01 April 2008

M. Perez-Camps  and
F. Garcia-Ximenez
M. Perez-Camps*
Affiliation:
Laboratory of Animal Reproduction and Biotechnology (LARB-UPV), Polytechnic University of Valencia, Camino de Vera 14, 46071 Valencia, Spain
*
E-mail: [email protected]
Get access

Abstract

With the aim of carrying out chimaerism and somatic cell–midblastula transition (MBT) embryos co-culture experiments in freshwater fish species, we evaluated the effect of osmolarity and composition of two media commonly used in cell fish culture on MBT zebrafish embryos and their further development and survival. To this end, wild zebrafish dechorionated embryos in midblastula stage were cultured for 6 days (Experiment 1: 189 embryos) or 1 h (Experiment 2: 150 embryos) in three different media: Hanks’ 10% (H-10), 35 mOsm; Hanks’ 100% (CH), 315 mOsm; and L-15 with serum (L-15: 315 mOsm). High osmolarity affected the survival rate (6 days: L-15: 45.1% v. CH: 72.34% v. H-10: 100%, P < 0.05; after 6 days: 0% both in L-15 and CH) and slowed their developmental timing. Embryos showed tail deformation (curly) as well as body paralysis at 48 h when they showed tail movements at 28 h. Differences in tail deformation were observed between high-osmolarity groups (CH: 85.10% v. L-15: 98.04%; P < 0.05). In Experiment 2, no effects on survival rate were observed. Teratogenic effects were only observed in L-15 (L-15: 12.98% v. CH: 0%; P< 0.05). Loss of motility was not detected in any group at 48 h. Optimum osmolar condition for cultured cells and also embryonic cells is around 315 mOsm and so, during chimaerism experiments (usually practised at MBT stage), present results indicate that midblastula embryos can acceptably bear the effects caused by 315 mOsm (CH) for 1 h, even though this involves a certain delay in developmental timing.

Keywords

hyperosmolaritychimaerismcell cultureembryozebrafish
Type
Full Paper
Information
animal , Volume 2 , Issue 4 , April 2008 , pp. 595 - 599
Copyright
Copyright © The Animal Consortium 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Adams, SL, Zhang, T, Rawson, DM 2005. The effect of external medium composition on membrane water permeability of zebrafish (Danio rerio) embryos. Theriogenology 64, 1591–1602.CrossRefGoogle ScholarPubMed
Boiani, M, Eckardt, S, Leu, NA, Schöler, HR, McLaughlin, KJ 2003. Pluripotency deficit in clones overcome by clone-clone aggregation: epigenetic complementation? EMBO 22, 5304–5312.CrossRefGoogle ScholarPubMed
Burnside, A, Collas, P 2002. Induction of Oct-3/4 expression in somatic cells by gap junction-mediated signalling from blastomeres. European Journal of Cell Biology 81, 585–591.CrossRefGoogle ScholarPubMed
Calvi, SL, Maisse, G 1998. Cryopreservation of rainbow trout (Oncorhynchus mykiss) blastomeres: influence of embryo stage on postthaw survival rate. Cryobiology 36, 225–262.CrossRefGoogle ScholarPubMed
Cervera, RP, García-Ximénez, F 2004a. Somatic cloning in rabbits: an element of reflection. In Animal Science Papers and Reports 22: Suppliment 1 (ed. JA Modlin′ski and J Karasiewicz), pp. 71–79. Institute of Genetics and Animal Breeding, Jastrzebiec, Poland.Google Scholar
Cervera, RP, García-Ximénez, F 2004b. Subzonal older adult fibroblast insertion in both in vivo-fertilized and nuclear transfer rabbit zygotes and embryos: effects on further in vitro embryo development. Cloning and Stem Cells 6, 315–326.CrossRefGoogle ScholarPubMed
Escribá, MJ, Silvestre, MA, Saeed, AM, García-Ximénez, F 2001. Comparison of the effect of two different handling media on rabbit zygote developmental ability. Reproduction, Nutrition, Development 41, 181–186.CrossRefGoogle ScholarPubMed
Gilmour, DT, Jessen, JR, Lin, S 2002. Manipulating gene expression in the zebrafish. In Zebrafish: A practical approach (ed. C Nüsslein-Volhard and R Dahm), pp. 121–143. Oxford University Press, New York.CrossRefGoogle Scholar
Haddy, JA, Pankhurst, NW 2000. The effects of salinity on reproductive development, plasma steroid levels, fertilisation and egg survival in black bream Acanthopagrus butcheri. Aquaculture 188, 115–131.CrossRefGoogle Scholar
Hart, PR, Purser, GJ 1995. Effects of salinity and temperature on eggs and yolk sac larvae of the greenback flounder (Rhombosolea tapirina Günther, 1862). Aquaculture 136, 221–230.CrossRefGoogle Scholar
Sawant, MS, Zhang, S, Li, L 2001. Effect of Salinity on development of zebrafish (Brachidanio rerio). Current Science 81, 1347–1350.Google Scholar
Sekirina, GG, Neganova, IE 1995. The microenvironment created by non blocking embryos in aggregates may rescue blocking embryos via cell-embryo adherent contacts. Zygote 3, 313–324.CrossRefGoogle ScholarPubMed
Van Der Wal, EJ 1985. Effects of temperature and salinity on the hatch rate and survival of Australian bass (Macquaria novemaculeata) eggs and yolk-sac larvae. Aquaculture 47, 239–244.CrossRefGoogle Scholar
Westerfield, M 2003. The zebrafish book: a guide for the laboratory use of zebrafish (Brachydanio rerio). Oregon University, Eugene, Oregon.Google Scholar
Young, PS, Dueñas, CE 1993. Salinity tolerance of fertilized eggs and yolk-sac larvae of the rabbitfish Siganus guttatus (Bloch). Aquaculture 112, 363–377.CrossRefGoogle Scholar