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7. add fish: the goldfish should be kept with or without a saltwater environment. if they are in a saltwater environment, they may need to be acclimated to the water. for this purpose, it’s best to leave them in the tank for a few days before moving them to a new tank. 8. add the fish: while the fish are acclimating, you can add the goldfish to the tank. add the fish slowly, a few at a time, and watch to ensure the water is even.expression of the chordin gene was analyzed by rna in situ hybridization using a digoxigenin-labeled crna probe that spanned the entire coding region of the chordin gene ( fig. s2a-f ). in a wild-type embryo, expression of the chordin gene was detected in the presomitic mesoderm, notochord, floor plate of the hindbrain and ventral spinal cord ( fig. s2c and s2d ). in goldfish with reduced chordin gene expression, notochord, hindbrain and ventral spinal cord expression of the chordin gene was markedly reduced ( fig. s2e-f ). these findings are consistent with the embryonic phenotypes (wild-type, weakly-ventralized, bifurcated fin fold and severely-ventralized). the twin-tail morphology of goldfish was reproduced by knockdown of the chordin gene in zebrafish ( fig. s2h and s2i ); however, this mutation did not affect the expression of the chordin gene in zebrafish. these data suggest that the role of the chordin gene in goldfish and zebrafish is different.in order to confirm that the twin-tail phenotype was reproduced by deficiency of the chordin gene, we analyzed the mrna expression of a series of genes that are known to be involved in tail patterning in zebrafish ( fig. 2a-d ). as expected, knockdown of the chordin gene in zebrafish resulted in drastic changes in gene expression levels: the gsc gene was down-regulated, whereas msgn was up-regulated in the single-tail zebrafish ( fig. 2b and 2d ). in addition, the expression levels of the chda and -b genes in goldfish were decreased ( fig. 2a and 2c ). these expression data indicate that the chordin gene is involved in tail development in both zebrafish and goldfish.
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