Molecular cloning, gene expression in albino mutants and gene knockdown studies of tyrosinase mRNA in rainbow trout

Pigment Cell Research
Surintorn BoonanuntanasarnToshio Takeuchi

Abstract

Tyrosinase has a role in melanin synthesis and several defects of the tyrosinase gene lead to albinism. Here, we cloned and characterized rainbow trout tyrosinase cDNAs and carried out the molecular and biochemical characterization of albino mutants. Two types of cDNA were cloned: tyrosinase-1 (Tyr-1) and tyrosinase-2 (Tyr-2). Both contained regions predicted to encode structural features of tyrosinase, and phylogenetic analysis confirmed that Tyr-1 and Tyr-2 were members of the tyrosinase family. Tyr-1 transcripts were first detected in embryos at 5 d post-fertilization (dpf) and Tyr-2 transcripts at 15 dpf. 3,4-dihydroxyphenylalanine assays revealed significantly reduced tyrosinase activities in dominant and recessive albino mutants compared with wild-type embryos. However, reverse-transcription PCR showed no differences in the amounts or lengths of the coding regions of Tyr-1 and Tyr-2 transcripts between wild-type embryos and albino mutants. Antisense morpholino oligonucleotides (AMOs) designed to knockdown tyrosinase gene expression in wild-type embryos led to reduced pigmentation in the retina and skin of embryos at 25 and 35 dpf, respectively. Furthermore, the tyrosinase activities of AMO-treated embryos were significant...Continue Reading

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Citations

Jan 16, 2013·Proceedings of the National Academy of Sciences of the United States of America·Seungki LeeGoro Yoshizaki
Jan 13, 2006·The Journal of Heredity·J M BlancEdwige Quillet
Apr 17, 2009·Genome Génome / Conseil National De Recherches Canada·K Ocalewicz, S Dobosz
May 15, 2007·BMC Evolutionary Biology·Ingo BraaschJean-Nicolas Volff
Jun 15, 2010·Comparative Biochemistry and Physiology. Part D, Genomics & Proteomics·Goro YoshizakiYutaka Takeuchi
Sep 13, 2005·Biochemical and Biophysical Research Communications·Surintorn BoonanuntanasarnGoro Yoshizaki
Sep 2, 2016·Biochemical and Biophysical Research Communications·Seungki LeeGoro Yoshizaki
May 31, 2014·Biology of Reproduction·Makoto HayashiGoro Yoshizaki
Nov 3, 2015·Scientific Reports·Seungki LeeGoro Yoshizaki
Dec 15, 2006·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Jennifer M FitzpatrickKarl F Hoffmann
Mar 13, 2010·Development·Goro YoshizakiTomoyuki Okutsu

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