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Overcoming a permeability barrier by microinjecting cryoprotectants into zebrafish embryos (Brachydanio rerio).

Author(s) : JANIK M., KLEINHANS F. W., HAGEDORN M.

Type of article: Article

Summary

The goal of this research was to examine the developmental effects on zebrafish embryos (Brachydanio rerio) when cryoprotectants were directly microinjected into the yolk. The objectives were to: 1) determine the final concentration of propylene glycol (PG) and dimethyl sulfoxide (DMSO) that the embryos could tolerate without causing teratogenic effects; 2) determine if the toxicity of DMSO could be reduced by the simultaneous use of various proportions of amides; and 3) examine whether this intracellular cryoprotectant incorporation could reduce the cryodamage to the yolk syncytial layer (YSL) after vitrification. The aim was to overcome the permeability barrier of the YSL. Intracellular PG produced better survival than DMSO, (P < 0.05). Embryos tolerated both 10- and 30-nl microinjections of PG, yielding final concentrations of 2.3 and 5.0 M within the yolk, resulting in 70 plus or minus 3 and 35 plus or minus 4% survival at day 5, respectively. With DMSO, survival was lower than PG at 60 plus or minus 4 and 14 plus or minus 4% at 2.4 and 5.2 M. Unlike other cellular systems, the presence of amides, specifically acetamide or formamide, did not reduce the toxicity of DMSO in zebrafish embryos (P > 0.05). Vitrification trials showed a 25% dehydration of the yolk, yielding an effective PG concentration of 5.9 M. The incorporation of this vitrifiable concentration of PG was not sufficient to improve the postthaw morphology of the YSL (P > 0.05).

Details

  • Original title: Overcoming a permeability barrier by microinjecting cryoprotectants into zebrafish embryos (Brachydanio rerio).
  • Record ID : 2001-2952
  • Languages: English
  • Source: Cryobiology - vol. 41 - n. 1
  • Publication date: 2000/08

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