PUBLICATION

Characterization of zebrafish mutants with defects in bone calcification during development

Authors
Xi, Y., Chen, D., Sun, L., Li, Y., and Li, L.
ID
ZDB-PUB-131010-6
Date
2013
Source
Biochemical and Biophysical Research Communications   440(1): 132-136 (Journal)
Registered Authors
Li, Lei, Li, Yuhao
Keywords
zebrafish, mutant, bone development, calcification
MeSH Terms
  • Animals
  • Bone Development
  • Calcification, Physiologic*
  • Cartilage/embryology
  • Cartilage/metabolism
  • Female
  • Gene Expression Regulation, Developmental
  • Male
  • Mutation*
  • Spine/abnormalities*
  • Spine/embryology*
  • Spine/metabolism
  • Zebrafish/embryology*
  • Zebrafish/genetics*
PubMed
24051095 Full text @ Biochem. Biophys. Res. Commun.
Abstract

Using the fluorescent dyes calcein and alcian blue, we stained the F3 generation of chemically (ENU) mutagenized zebrafish embryos and larvae, and screened for mutants with defects in bone development. We identified a mutant line, bone calcification slow (bcs), which showed delayed axial vertebra calcification during development. Before 4–5 days post-fertilization (dpf), the bcs embryos did not display obvious abnormalities in bone development (i.e., normal number, size and shape of cartilage and vertebrae). At 5–6 dpf, when vertebrae calcification starts, bcs embryos began to show defects. At 7 dpf, for example, in most of the bcs embryos examined, calcein staining revealed no signals of vertebrae mineralization, whereas during the same developmental stages, 2–14 mineralized vertebrae were observed in wild-type animals. Decreases in the number of calcified vertebrae were also observed in bcs mutants when examined at 9 and 11 dpf, respectively. Interestingly, by 13 dpf the defects in bcs mutants were no longer evident. There were no significant differences in the number of calcified vertebrae between wild-type and mutant animals. We examined the expression of bone development marker genes (e.g., Sox9b, Bmp2b, and Cyp26b1, which play important roles in bone formation and calcification). In mutant fish, we observed slight increases in Sox9b expression, no alterations in Bmp2b expression, but significant increases in Cyp26b1 expression. Together, the data suggest that bcs delays axial skeletal calcification, but does not affect bone formation and maturation.

Genes / Markers
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Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping