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Spermidine, but not spermine, is essential for pigment pattern formation in zebrafish.

Frohnhöfer HG, Geiger-Rudolph S, Pattky M, Meixner M, Huhn C, Maischein HM, Geisler R, Gehring I, Maderspacher F, Nüsslein-Volhard C, Irion U - Biol Open (2016)

Bottom Line: Here we identify idefix, a mutation in the zebrafish gene encoding the enzyme spermidine synthase, leading to a severe reduction in spermidine levels as shown by capillary electrophoresis-mass spectrometry.This allows us to uncouple them from events occurring later during colour patterning.Thus, zebrafish provide a vertebrate model to study the in vivo effects of polyamines.

View Article: PubMed Central - PubMed

Affiliation: Max-Planck-Institut für Entwicklungsbiologie, Abteilung 3, Spemannstrasse 35, Tübingen 72076, Germany.

No MeSH data available.


Related in: MedlinePlus

A knockout mutation in spermine synthase has no phenotypic consequences. (A) Chromatograms of wild-type, homozygous and heterozygous mutants showing that the CRISPR knockout of sms resulted in an 8 bp insertion. (B) Homozygous fish are viable and fertile and show no phenotype. Scale bar: 5 mm.
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BIO018721F7: A knockout mutation in spermine synthase has no phenotypic consequences. (A) Chromatograms of wild-type, homozygous and heterozygous mutants showing that the CRISPR knockout of sms resulted in an 8 bp insertion. (B) Homozygous fish are viable and fertile and show no phenotype. Scale bar: 5 mm.

Mentions: Spermidine is the substrate for the synthesis of the longer polyamine spermine. To investigate whether the ide phenotype is directly caused by the lack of spermidine or whether the absence of spermine is responsible, we generated knock out mutants for the gene encoding spermine synthase (sms). We used the CRISPR/Cas9 system to target exon 2 of the gene and recovered a mutant with an 8 bp insertion, leading to a frame shift resulting in a truncated protein of 50 amino acids followed by 20 unrelated residues and then a stop codon. Fish homozygous for this mutation are viable, fertile and normally pigmented (Fig. 7). This shows that it is the absence of spermidine that leads to the ide phenotype; spermine is not essential for zebrafish.Fig. 7.


Spermidine, but not spermine, is essential for pigment pattern formation in zebrafish.

Frohnhöfer HG, Geiger-Rudolph S, Pattky M, Meixner M, Huhn C, Maischein HM, Geisler R, Gehring I, Maderspacher F, Nüsslein-Volhard C, Irion U - Biol Open (2016)

A knockout mutation in spermine synthase has no phenotypic consequences. (A) Chromatograms of wild-type, homozygous and heterozygous mutants showing that the CRISPR knockout of sms resulted in an 8 bp insertion. (B) Homozygous fish are viable and fertile and show no phenotype. Scale bar: 5 mm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920196&req=5

BIO018721F7: A knockout mutation in spermine synthase has no phenotypic consequences. (A) Chromatograms of wild-type, homozygous and heterozygous mutants showing that the CRISPR knockout of sms resulted in an 8 bp insertion. (B) Homozygous fish are viable and fertile and show no phenotype. Scale bar: 5 mm.
Mentions: Spermidine is the substrate for the synthesis of the longer polyamine spermine. To investigate whether the ide phenotype is directly caused by the lack of spermidine or whether the absence of spermine is responsible, we generated knock out mutants for the gene encoding spermine synthase (sms). We used the CRISPR/Cas9 system to target exon 2 of the gene and recovered a mutant with an 8 bp insertion, leading to a frame shift resulting in a truncated protein of 50 amino acids followed by 20 unrelated residues and then a stop codon. Fish homozygous for this mutation are viable, fertile and normally pigmented (Fig. 7). This shows that it is the absence of spermidine that leads to the ide phenotype; spermine is not essential for zebrafish.Fig. 7.

Bottom Line: Here we identify idefix, a mutation in the zebrafish gene encoding the enzyme spermidine synthase, leading to a severe reduction in spermidine levels as shown by capillary electrophoresis-mass spectrometry.This allows us to uncouple them from events occurring later during colour patterning.Thus, zebrafish provide a vertebrate model to study the in vivo effects of polyamines.

View Article: PubMed Central - PubMed

Affiliation: Max-Planck-Institut für Entwicklungsbiologie, Abteilung 3, Spemannstrasse 35, Tübingen 72076, Germany.

No MeSH data available.


Related in: MedlinePlus