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Morpholino oligonucleotide knockdown reveals extracellular calcium‐sensing receptor role in zebrafish skeletal development
Author(s) -
Herberger Amanda L,
Loretz Christopher A
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.937.19
Subject(s) - gene knockdown , zebrafish , morpholino , biology , ciona intestinalis , microbiology and biotechnology , vertebrate , calcium sensing receptor , parathyroid hormone , calcium , medicine , genetics , gene
The complex vertebrate skeleton depends on regulated cell activities to lay down protein matrix and mineral components. Bone, a distinctive vertebrate characteristic, is a storage site for physiologically‐important calcium ion. The extracellular calcium‐sensing receptor (CaSR), a G protein‐coupled receptor that is tightly linked to homeostatic regulation of calcium by endocrine glands, ion‐transporting epithelia and bone, is restricted to the chordate‐vertebrate evolutionary lineage. Our overall research objective is to understand the functional linkage of CaSRs to skeleton in the context of vertebrate evolution in diverse habitats. Since little is known about CaSR in skeletal biology of non‐mammalian vertebrates, immunohistochemistry was applied to adult and embryonic zebrafish, revealing CaSR protein expression in many tissues, including chondrocytes and notochordal sheath cells. Morpholino oligonucleotide (MO) knockdown was used to discover CaSR role(s) in this zebrafish model system. By RT‐PCR, MO treatment reduced CaSR mRNA expression measured at 1 and 2 dpf in TU strain fish. Corresponding to the MO knockdown of normally‐spliced CaSR mRNA, we observed a phenotype characterized by stunted growth and disorganization of the notochordal component of axial skeleton by 1 dpf. We conclude that, like to its essential role in bone development in mammals, CaSR is important in skeletogenesis in fish.