Abstract
As a new type of AFPs, AFPIV has been firstly identified in longhorn sculpin (Myoxocephalus octodecimspinosus), and in recent years, its cDNA and amino acid sequence have been reported, and its pancreatic synthesis has been firstly reported in polar fish. However, its expression patterns during fish embryogenesis have not been elucidated yet. By differential screening, we cloned the CagAFPIV in gibel carp, Carassius auratus gibelio, demonstrated its predominant expression during embryogenesis. RT-PCR detection revealed that CagAFPIV was first transcribed from blastula stage and kept a high level during embryogenesis and declined remarkably in hatched larva. In situ hybridization revealed that CagAFPIV transcripts were firstly distributed over the margin and marginal blastomere in blastula stage embryos, at the early-gastrula stage the positive signals distributed in the marginal cells and the internalization cells, and later restricted to the cells the yolk syncytial layer (YSL) from later gastrula stage to larva stage. Consistently, the CagAFPIV protein also kept a high level during embryogenesis, and the high protein level retained some days after the larva hatched. Our work, for the first time, revealed the dynamic expression and distribution of CagAFPIV during embryogenesis.
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Abbreviations
- AFPIV:
-
Antifreeze type IV protein
- CagAFPIV :
-
Carassius auratus gibelio antifreeze type IV protein
- CagApo-14 :
-
Carassius auratus gibelio Apo-14
- Cagfetuin-B :
-
Carassius auratus gibelio fetuin-B
- nt:
-
Nucleotide
- ORF:
-
Open reading frame
- UTR:
-
Untranslated region
- SMART:
-
Switch mechanism at the 5′ end of RNA templates
- PBS:
-
Phosphate saline buffer
- FITC:
-
Fluorescein isothiocyanate
- HE:
-
Hematoxylin and Eosin
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Acknowledgments
This work was supported by grants from the National Major Basic Research Program (2004CB117401) and the National Natural Science Foundation of China—Youth Foundation (grant No. 30700440).
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The nucleotide sequence data reported in this paper have been submitted to the GenBank under accession number: CagAFPIV (AY365004).
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Liu, JX., Zhai, YH. & Gui, JF. Molecular characterization and expression pattern of AFPIV during embryogenesis in gibel carp(Carassiu auratus gibelio). Mol Biol Rep 36, 2011–2018 (2009). https://doi.org/10.1007/s11033-008-9412-3
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DOI: https://doi.org/10.1007/s11033-008-9412-3