ECTS2016 Poster Presentations Cell biology: osteoclasts and bone resorption (35 abstracts)
Identification of G protein-coupled receptor 137B (GPR137b) function in mouse and zebrafish osteoclasts
Katia Urso 1, , Joana Caetano-Lopes 2, , Pui Lee 1, , Katrin Henke 2, , Meera Sury 1, , Jeff Duryea 1, , Peter Nigrovic 1, , Antonios Aliprantis 1, , Matt Harris 2, & Julia Charles 1,
1Brigham and Womne’s Hospital, Boston, Massachusetts, USA, 2Harvard Medical School, Boston, Massachusetts, USA, 3Boston Children’s Hospital, Boston, Massachusetts, USA.
Analysis of genome wide data such as transcriptomics can identify genes of potential interest to bone biology. These techniques are primarily hypothesis-generating. Determining the role of candidate factors in bone ultimately requires in vivo experiments. Gene expression analysis of mouse osteoclast differentiation identified G protein-coupled receptor 137B (GPR137b) as highly upregulated. GPR137b is an orphan seven-pass transmembrane receptor of unknown function. We demonstrated co-localization of GPR137b and TRAP in osteoclasts on histologic sections from mouse femurs. To study GPR137b function in vitro we used the CRISPR/Cas9 system to generate knock-out murine osteoclast precursor cell lines. Gpr137b−/− osteoclasts cultured on mineralized matrix shown increased bone resorbtion activity. To determine whether GPR137b is important for osteoclast function in vivo, we used CRISPR/Cas9 to specifically knock-out the orthologous gene in zebrafish. Gpr137b homozygous mutants are viable and fertile and do not display overt morphological defects as adults. However, analysis of pits formation on scales of gpr137b−/− zebrafish demonstrated an increase in bone resorption, results in line with the data generated with Gpr137b−/− mouse cells. These data suggest a role for GPR137b as a negative regulator of osteoclast activity. To determine if the increase in bone resorption in gpr137−/− osteoclasts alters skeletogenesis in zebrafish, we are utilizing a robust microCT method we developed to measure and evaluate bone mass and shape. To further study the role of GPR137b in osteoclast function we are assessing osteoclast behavior in vivo using a cathepsinK-dsRed reporter line in WT and mutant zebrafish.
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Bone Abstracts
ISSN 2052-1219 (online)
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