Spatial expression patterns of skin-type antifreeze protein in winter flounder (Pseudopleuronectes americanus) epidermis following metamorphosis

2003 ◽  
Vol 257 (1) ◽  
pp. 78-86 ◽  
Author(s):  
Harry M. Murray ◽  
Choy L. Hew ◽  
Garth L. Fletcher
Keyword(s):  
Expression Patterns ◽  
Antifreeze Protein ◽  
Winter Flounder ◽  
Skin Type ◽  
Pseudopleuronectes Americanus ◽  
Spatial Expression

Evidence for Antifreeze Protein Gene Transfer in Atlantic Salmon (Salmo salar)

1988 ◽  
Vol 45 (2) ◽  
pp. 352-357 ◽  
Author(s):  
Garth L. Fletcher ◽  
Margaret A. Shears ◽  
Madonna J. King ◽  
Peter L. Davies ◽  
Choy L. Hew
Keyword(s):  
Atlantic Salmon ◽  
Salmo Salar ◽  
Cold Water ◽  
Restriction Enzymes ◽  
Antifreeze Protein ◽  
Winter Flounder ◽  
Control Fish ◽  
Pseudopleuronectes Americanus ◽  
Atlantic Salmon Salmo Salar ◽  
Antifreeze Polypeptides

Atlantic salmon (Salmo salar) freeze to death if they come into contact with ice at water temperatures below −0.7 °C. Consequently, sea-pen culture of this species in cold water is severely limited. Winter flounder (Pseudopleuronectes americanus) survive in ice-laden seawater by producing a set of antifreeze polypeptides (AFP). We are attempting to make the Atlantic salmon more freeze resistant by transferring antifreeze protein genes from the winter flounder to the genome of the salmon. Salmon eggs were microinjected with linearized DNA after fertilization. Individual fingerlings (1–2 g) were analyzed for flounder AFP genes by genomic Southern blotting. DNA from 2 out of 30 fingerlings showed hybridization to the flounder DNA probe. Hybridization bands following cleavage by restriction enzymes Sst l and Bam HI were identical to those of the injected DNA. Hybridization following Hind III digestion indicated that the flounder AFP gene was linked to the salmon genome. These hybridization signals were absent in the DNA from control fish. The intensity of the hybridization signals indicated that there was on average at least one copy of the AFP gene present per cell.

scholarly journals Expression of genes involved in key metabolic processes during winter flounder (Pseudopleuronectes americanus) metamorphosis

2013 ◽  
Vol 91 (3) ◽  
pp. 156-163 ◽  
Author(s):  
Marie Vagner ◽  
Benjamin de Montgolfier ◽  
Jean-Marie Sévigny ◽  
Réjean Tremblay ◽  
Céline Audet
Keyword(s):  
Gene Expression ◽  
Expression Patterns ◽  
Pentose Phosphate ◽  
Winter Flounder ◽  
Benthic Habitat ◽  
Pseudopleuronectes Americanus ◽  
Metabolic Processes ◽  
Expression Of Genes ◽  
Molecular Events ◽  
Gh Gene

The aim of this study was to better understand the molecular events governing ontogeny in winter flounder (Pseudopleuronectes americanus (Walbaum, 1792)). The expression of seven genes involved in key metabolic processes during metamorphosis were measured at settlement (S0), at 15 (S15), and 30 (S30) days after settlement and compared with those in pelagic larvae prior to settlement (PL). Two critical stages were identified: (1) larval transit from the pelagic to the benthic habitat (from PL to S0) and (2) metamorphosis maturation, when the larvae stay settled without growth (from S0 to S30). Growth hormone (gh) gene expression significantly increased at S0. At S30, an increase in cytochrome oxidase (cox) gene expression occurred with a second surge of gh gene expression, suggesting that enhanced aerobic capacity was supporting growth before the temperature decrease in the fall. Expression patterns of pyruvate kinase (pk), glucose-6-phosphate dehydrogenase (g6pd), and bile salt activated lipase (bal) genes indicated that energy synthesis may be mainly supplied through glycolysis in PL, through the pentose–phosphate pathway at settlement, and through lipid metabolism at S30. The expression of the heat-shock protein 70 (hsp70), superoxide dismutase (sod), cox, and peroxiredoxin-6 (prx6) genes revealed that oxidative stress and the consequent development of antioxidative protection were limited during the PL stage, reinforced at settlement, and very high at S30, certainly owing to the higher growth rate observed at this period.

Tissue distribution of fish antifreeze protein mRNAs

1992 ◽  
Vol 70 (4) ◽  
pp. 810-814 ◽  
Author(s):  
Zhiyuan Gong ◽  
Garth L. Fletcher ◽  
Choy L. Hew
Keyword(s):  
Northern Blot Analysis ◽  
Antifreeze Protein ◽  
The Body ◽  
Winter Flounder ◽  
Type I ◽  
Pseudopleuronectes Americanus ◽  
Sea Raven ◽  
Hemitripterus Americanus ◽  
Afp Mrna ◽  
Distribution Of Fish

The presence of fish antifreeze protein (AFP) mRNA was examined in a variety of tissues from the winter flounder (Pseudopleuronectes americanus), sea raven (Hemitripterus americanus), and ocean pout (Macrozoarces americanus), each of which contains one of the three known AFP types. Northern blot analysis indicates that whereas the AFP mRNA is restricted to liver in sea raven (type II AFP), significant amounts of mRNA are present in many other tissues in both winter flounder (type I) and ocean pout (type III). These results indicate that in sea raven, antifreeze protein synthesis only occurs in the liver, whereas in the ocean pout and winter flounder, synthesis occurs in many tissues throughout the body. These investigations are relevant to understanding the mode of action of these polypeptides.

Antifreeze peptides confer freezing resistance to fish

1986 ◽  
Vol 64 (9) ◽  
pp. 1897-1901 ◽  
Author(s):  
Garth L. Fletcher ◽  
Ming H. Kao ◽  
Ron M. Fourney
Keyword(s):  
Seasonal Changes ◽  
Antifreeze Proteins ◽  
Antifreeze Protein ◽  
Winter Flounder ◽  
Salmo Gairdneri ◽  
Pseudopleuronectes Americanus ◽  
Freezing Resistance ◽  
Protein Levels ◽  
Protein Receptors ◽  
Species Specific

It has been widely accepted that plasma antifreeze proteins are directly responsible for the ability of many marine teleosts to survive in ice-laden seawater. However, there appears to be no direct experimental evidence to indicate that this assumption is correct. In the present study winter flounder (Pseudopleuronectes americanus) showed seasonal changes in freezing resistance that were quantitatively the same as the seasonal changes in plasma antifreeze protein levels. Moreover, when winter flounder antifreeze proteins were injected into rainbow trout (Salmo gairdneri) (a species that does not normally possess antifreeze proteins) they increased the freezing resistance of the trout in direct proportion to plasma antifreeze protein levels attained. These studies indicate that antifreeze proteins are directly responsible for the ability of many marine teleosts to survive icy seawater at temperatures below the colligative freezing points of their blood. There appears to be no requirement for species-specific antifreeze protein receptors in the fish in order for them to act.

Localization of cells from the winter flounder gill expressing a skin type antifreeze protein gene

2002 ◽  
Vol 80 (1) ◽  
pp. 110-119 ◽  
Author(s):  
Harry M Murray ◽  
Choy L Hew ◽  
Ken R Kao ◽  
Garth L Fletcher
Keyword(s):  
In Situ Hybridization ◽  
Antifreeze Protein ◽  
Tissue Expression ◽  
Cross Reactivity ◽  
Winter Flounder ◽  
Skin Type ◽  
General Distribution ◽  
Pavement Cells ◽  
Peripheral Tissues

In situ hybridization on whole mounts and paraffin-sectioned winter flounder (Pleuronectes americanus) gill, using riboprobes specific to a skin type antifreeze protein (AFP) gene, showed a mRNA distribution associated with cells throughout the filament and the lamellae. Immunohistochemistry using antibodies for a skin-type AFP identified cells corresponding to those detected using in situ hybridization. Parallel experiments with antibodies for chloride-cell markers showed that these cells were not involved in antifreeze-protein expression. Similarly, goblet cells did not show cross-reactivity with the AFP antibodies. This general distribution suggested that pavement cells were likely involved. Reverse transcription polymerase chain reaction using gill cDNA templates and subsequent sequencing of the products confirmed the presence of skin type AFP transcripts in this tissue. Expression of a AFP in this area may act as a first line of defence against ice-crystal migration into peripheral tissues.

Cloning, eukaryotic expression and spatial expression patterns of porcine MNSFβ

2013 ◽  
Vol 35 (12) ◽  
pp. 1377-1383
Author(s):  
Jing-Na WANG ◽  
Peng-Fei JIANG ◽  
Zhan-Zhan KANG ◽  
Deng-Yun LI ◽  
Lin-Lin HUA ◽  
...  
Keyword(s):  
Expression Patterns ◽  
Eukaryotic Expression ◽  
Spatial Expression
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