PUBLICATION

Characterization of biklf/klf17-deficient zebrafish in posterior lateral line neuromast and hatching gland development

Authors
Suzuki, H., Ishizaka, T., Yanagi, K., Sone, R., Sunaga, Y., Ohga, R., Kawahara, A.
ID
ZDB-PUB-190929-3
Date
2019
Source
Scientific Reports   9: 13680 (Journal)
Registered Authors
Kawahara, Atsuo, Suzuki, Hiroaki
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Hematopoiesis/genetics
  • Kruppel-Like Transcription Factors/deficiency*
  • Kruppel-Like Transcription Factors/genetics*
  • Lateral Line System/embryology*
  • Lateral Line System/metabolism
  • Zebrafish/embryology*
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zebrafish Proteins/deficiency*
  • Zebrafish Proteins/genetics*
(all 14)
PubMed
31558744 Full text @ Sci. Rep.
Abstract
Krüpple-like factors (Klfs) are highly conserved zinc-finger transcription factors that regulate various developmental processes, such as haematopoiesis and cardiovascular development. In zebrafish, transient knockdown analysis of biklf/klf17 using antisense morpholino suggests the involvement of biklf/klf17 in primitive erythropoiesis and hatching gland development; however, the continuous physiological importance of klf17 remains uncharacterized under the genetic ablation of the klf17 gene among vertebrates. We established the klf17-disrupted zebrafish lines using the CRISPR/Cas9 technology and performed phenotypic analysis throughout early embryogenesis. We found that the klf17-deficient embryos exhibited abnormal lateral line neuromast deposition, whereas the production of primitive erythrocytes and haemoglobin production were observed in the klf17-deficient embryos. The expression of lateral line neuromast genes, klf17 and s100t, in the klf17-deficient embryos was detected in posterior lateral line neuromasts abnormally positioned at short intervals. Furthermore, the klf17-deficient embryos failed to hatch and died without hatching around 15 days post-fertilization (dpf), whereas the dechorionated klf17-deficient embryos and wild-type embryos were alive at 15 dpf. The klf17-deficient embryos abolished hatching gland cells and Ctsl1b protein expression, and eliminated the expression of polster and hatching gland marker genes, he1.1, ctsl1b and cd63. Thus, the klf17 gene plays important roles in posterior lateral line neuromast and hatching gland development.
Genes / Markers
Marker Marker Type Name
cd63GENECD63 molecule
ctslbGENEcathepsin Lb
gata1aGENEGATA binding protein 1a
hbbe3GENEhemoglobin beta embryonic-3
he1.1GENEhatching enzyme 1, tandem duplicate 1
klf17GENEKruppel like factor 17
lyzGENElysozyme
s100tGENES100 calcium binding protein T
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Figures
Figure Gallery (7 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
WT + MO1-klf17standard conditions26+ somites
klf17uy21/+; klf17uy22/+standard conditionsPrim-5
klf17uy21/+; klf17uy22/+standard conditionsPrim-5
klf17uy21/+; klf17uy22/+standard conditionsPrim-5
klf17uy21/+; klf17uy22/+standard conditionsPrim-5
klf17uy21/+; klf17uy22/+standard conditionsLong-pec
klf17uy21/+; klf17uy22/+standard conditionsProtruding-mouth
klf17uy21/+; klf17uy22/+standard conditionsDay 6
klf17uy21/+; klf17uy22/+standard conditionsDay 6
klf17uy21/+; klf17uy22/+standard conditionsDays 7-13
1 - 10 of 25
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
uy21
    		Indel
    uy22
      		Indel
      uy23
        		Indel
        1 - 3 of 3
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        Human Disease / Model
        No data available
        Sequence Targeting Reagents
        Target Reagent Reagent Type
        klf17CRISPR1-klf17CRISPR
        klf17CRISPR2-klf17CRISPR
        klf17MO1-klf17MRPHLNO
        1 - 3 of 3
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        Fish
        Fish
        klf17uy21/+
        klf17uy21/+; klf17uy22/+
        klf17uy21/uy21
        klf17uy22/+
        klf17uy22/+; klf17uy23/+
        klf17uy22/uy22
        klf17uy23/+
        klf17uy23/uy23
        WT
        WT + MO1-klf17
        1 - 10 of 10
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        Antibodies
        No data available
        Orthology
        No data available
        Engineered Foreign Genes
        No data available
        Mapping
        No data available
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        Citation
        Suzuki, H., Ishizaka, T., Yanagi, K., Sone, R., Sunaga, Y., Ohga, R., Kawahara, A. (2019) Characterization of biklf/klf17-deficient zebrafish in posterior lateral line neuromast and hatching gland development. Scientific Reports. 9:13680.