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Published on: October 19, 2013
[Expression pattern of hoxd3 gene during early development of wild-type zebrafish embryos]
Li-ping Shu1, Zhi-xu He, Dong-jing Yao
1Tissue Engineering and Stem Cell Research Center, Guiyang Medical University, Guiyang 550004, China. gyslp456@yahoo.com.cn
Objective:
To investigate the expression pattern of hoxd3 gene during early embryogenesis and angiogenesis of wild-type zebrafish.
Methods:
Total RNA was extracted from embryos of zebrafish in different development stages by trizol. The cDNA of hoxd3 gene was amplified by RT-PCR. The RT-PCR product was ligated to pCS(2+) vector by T4 DNA ligatase polymerase and sequenced. T3 RNA polymerase in vitro transcription system was used to obtain the probe of digoxin-labeled anti-sense mRNA of hoxd3 gene. The expression pattern of hoxd3 was detected by whole embryo in situ hybridization (WISH) with anti-sense mRNA probe.
Results:
pCS(2+)-hoxd3 plasmid was successfully constructed, which was used to prepare anti-sense mRNA probe of hoxd3 in vitro. Expression pattern of hoxd3 gene was detected by WISH during zebrafish early embryogenesis and angiogenesis. It was observed that hoxd3 mRNA was expressed at the junction region of midbrain and hindbrain in wild-type zebrafish in embryos at 24 ≊72h postfertilization(hpf).
Conclusion:
hoxd3 gene is mainly expressed in nervous system of wide-type zebrafish embryos.
