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Loss of  L -gulonolactone oxidase ( GULO ), which catalyzes the last step of the ascorbic acid (AA) biosynthesis pathway, results in a complete lack of AA in several Osteichthyes fish species, including zebrafish. In this study,  sGULO , the active  GULO  gene from cloudy catshark ( Scyliorhinus torazame ) was cloned into zebrafish using the Gateway cloning method. The resulting  Tg(b-actin:sGULO:mCherry)  fish were analyzed for the effects of a reestablished AA pathway. Fluorescent microscopy and PCR were used to analyze the integration of the construct into the zebrafish genome. Catalytic activity of sGULO, AA production, growth-related characteristics, and gene expression were investigated to evaluate the effects of AA production in  Tg  fish. The mCherry fluorescent protein indicated the proper integration and expression of the  sGULO  construct in zebrafish. The  sGULO  gene was ubiquitously expressed in all the studied tissues and the enzyme activity indicated an increased AA production in  Tg  fish. The growth of  Tg  fish was also increased, and antioxidant system analysis suggests that reactive oxygen species production was reduced in  Tg  fish compared with wild type. Expression of the AA transporter  slc23a1  was significantly downregulated in  Tg  homozygous fish. These results collectively indicate the effects of reestablished AA synthesis in zebrafish.

frontiers in physiology

Growth and Antioxidant-Related Effects of the Reestablished Ascorbic Acid Pathway in Zebrafish (Danio rerio) by Genomic Integration of L-Gulonolactone Oxidase From Cloudy Catshark (Scyliorhinus torazame)