EurekaMag
+ Translate
+ Most Popular
Investigations on the organic drift in North Swedish streams
Limitations of Using Microsoft Excel Version 2016 (MS Excel 2016) for Statistical Analysis for Medical Research
Phloem necrosis of coffee in Surinam
Anatomy of Mystus seenghala IV Nervous system
Integrated farming system for gardenland conditions of Coimbatore district - an over view
Tetrapleure (Tetrapleura tetraptera), an unknown African medicinal and spice plant
The origin and phylogenetic significance of the trochophoran larvae 2. evolutionary significance of the larvae of coelomate worms and mollusks
Comparison of rice bran and maize bran as feeds for growing and fattening pigs
Enterobacter amnigenus. An unusual human pathogen
Influence of Seriboost foliar application on leaf yield and leaf protein content in mulberry (Morus spp.), in relation to silkworm cocoon production
The identity of the lipstick mold of cultivated mushrooms agaricus bisporus
Advantages and disadvantages of bordeaux mixture and of lime-sulphur used on apples in the growing season
'Pan-sukh' disease of Rice in the Central Provinces
Geological age of the Ptilophyllum flora; a critical reassessment
Study of vitellogenesis in birds; physiological phases & role of folliculin in vitellogenesis
Evaluation of WCT coconut and Komadan coconut
Therapy for acne with saccharomyces boulardii
Evidence for Late Cretaceous N-S dextral shear in the west-central crystalline core, North Cascades, Washington
'Rajeshwari' - a high-yielding white seeded variety of sesame for Andhra Pradesh
Manufacture of Ricotta cheese from whey fortified with skim milk powder using different acidulants
Occurrence of Eutrichophilus mexicanus (Rudow, 1866) and Eutrichophilus lobatus (Ewing, 1936) (Phthiraptera: Trichodectidae) on Sphiggurus villosus (Cuvier, 1825) (Rodentia: Erethizontidae) in Rio de
Factors affecting fruitfulness in durian (Durio zibethinus Murr.). I. Flowering and pollination
Bronchial cuff pressure change caused by left-sided double-lumen endobronchial tube displacement
Some Biological Applications of Organometallic Compounds
The composition of pampas-grass (Cortaderia argentea.)

Differential scanning calorimetry studies of intraembryonic freezing and cryoprotectant penetration in zebrafish (Danio rerio) embryos


Differential scanning calorimetry studies of intraembryonic freezing and cryoprotectant penetration in zebrafish (Danio rerio) embryos



Journal of Experimental Zoology 290(3): 299-310



ISSN/ISBN: 0022-104X

PMID: 11479909

DOI: 10.1002/jez.1060

Nucleation temperatures of intraembryonic water and cryoprotectant penetration in zebrafish embryos were studied using differential scanning calorimetry. The effects of embryo developmental stage, dechorionation, partial removal of yolk, cooling rate, and cryoprotectant treatment on the temperatures of intraembryonic freezing were investigated. Embryo stages were found to have a significant effect on the nucleation temperatures of intact embryos. Freeze onset temperatures of -11.9 +/- 1.5, -15.6 +/- 0.3, and -20.5 +/- 0.1 degrees C were obtained for intact embryos at 6-somite, prim-6, and high-pec stages, respectively. After dechorionation, the freeze onset temperatures of intraembryonic water shifted to significantly lower temperatures, being -23.5 +/- 0.8, -18.7 +/- 0.7, -24.9 +/- 0.8 degrees C for 6-somite, prim-6, and high-pec stages, respectively. Yolk-reduced high-pec stage embryos showed significantly lower nucleation temperatures with an average onset at -27.9 +/- 0.4 degrees C. The effect of cryoprotectant treatment on the nucleation temperatures of intraembryonic water varies among different embryo stages and different cryoprotectants. Thirty-minute treatment with 2 M methanol significantly decreased the nucleation temperatures of dechorionated 6-somite embryos whilst no temperature decrease was observed for prim-6 or yolk-reduced high-pec embryos. Thirty-minute exposure to 1 M propylene glycol did not significantly affect the nucleation temperatures of dechorionated 6-somite, prim-6, or yolk-reduced high-pec embryos. In order to increase the permeability of embryos to cryoprotectants, the yolk sacs of dechorionated embryos at 6-somite or prim-6 embryos were punctured with a sharp micro-needle before exposure to cryoprotectants. The punctured prim-6 embryos showed significantly lower temperatures of intraembryonic freezing after 30 min of exposure to 2 M methanol following the multi-punctures. The nucleation temperatures of punctured 6-somite or prim-6 embryos were also decreased significantly after exposure to 1 M propylene glycol for 30 min. These results suggested that in intact embryos, intraembryonic freezing appeared to be seeded by the external ice in the perivitelline fluid and that in dechorionated embryos (in the absence of external water) intraembryonic freezing was more likely a consequence of heterogeneous nucleation. Methanol was demonstrated to show a limited degree of penetration into prim-6 stage embryos, but it did not penetrate later-stage embryos such as prim-6 and yolk-reduced high-pec. No propylene glycol permeation was observed for embryos at all stages. However, multi-punctures of yolk resulted in the permeation of both cryoprotectants into prim-6 embryos and propylene glycol permeation into 6-somite embryos. These findings may have important implications in overcoming the problem associated with the low membrane permeability of zebrafish embryos to cryoprotectants.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 003713819

Download citation: RISBibTeXText

Related references

Short-term chilled storage of zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation. Zebrafish 12(1): 111-120, 2015

Studies on cryoprotectant toxicity to zebrafish (Danio rerio) oocytes. Cryo Letters 25(6): 415-424, 2004

Studies on cryoprotectant toxicity to zebrafish (Danio rerio) ovarian tissue fragments. Cryo Letters 32(1): 40-50, 2011

Studies on cryoprotectant toxicity to early stage zebrafish (Danio rerio) ovarian follicles. Cryo Letters 29(6): 477-483, 2008

Cryopreservation of zebrafish (Danio rerio) primordial germ cells by vitrification of yolk-intact and yolk-depleted embryos using various cryoprotectant solutions. Cryobiology: -, 2014

Cryopreservation of zebrafish (Danio rerio) primordial germ cells by vitrification of yolk-intact and yolk-depleted embryos using various cryoprotectant solutions. Cryobiology 67(3): 374-382, 2013

A differential scanning calorimetry dsc study of the freezing thawing behavior of cryoprotectant containing liposome dispersions. Acta Pharmaceutica Technologica 36(1): 17S, 1990

Ultrasound enhanced methanol penetration of zebrafish (Danio rerio) embryos measured by permittivity changes using impedance spectroscopy. European Biophysics Journal: Ebj 37(6): 1039-1044, 2008

Field emission scanning electron microscopy and transmission electron microscopy studies of the chorion, plasma membrane and syncytial layers of the gastrula-stage embryo of the zebrafish Brachydanio rerio : a consideration of the structural and functional relationships with respect to cryoprotectant penetration. Aquaculture Research 31(3): 325-336, 2000

Embryos of the zebrafish Danio rerio in studies of non-targeted effects of ionizing radiation. Cancer Letters 356(1): 91-104, 2015

Developmental stages of zebrafish (Danio rerio) embryos and toxicological studies using foldscope microscope. Cell Biology International 44(10): 1968-1980, 2020

Morpholino-mediated knockdown of ERα, ERβa, and ERβb mRNAs in zebrafish (Danio rerio) embryos reveals differential regulation of estrogen-inducible genes. Endocrinology 154(11): 4158-4169, 2013

Identification of differential transcript profiles between mutual crossbred embryos of zebrafish (Danio rerio) and Chinese rare minnow (Gobiocypris rarus) by cDNA-AFLP. Theriogenology 70(9): 1525-1535, 2008

Differential tissue-specific distribution of transcripts for the duplicated fatty acid-binding protein 10 (fabp10) genes in embryos, larvae and adult zebrafish (Danio rerio). Febs Journal 276(22): 6787-6797, 2009

Microplastic accumulation patterns and transfer of benzo[a]pyrene to adult zebrafish (Danio rerio) gills and zebrafish embryos. Environmental Pollution 235: 918-930, 2018