G protein‐coupled receptor gpr34l mutation affects thrombocyte function in zebrafish

• Published in: British journal of haematology 2018-02, Vol.180 (3), p.412-419
• Main Authors: Kim, Seongcheol, Alsrhani, Abdullah, Zafreen, Lala, Khandekar, Gauri, Marlow, Florence L, Abrams, Elliott W., Mullins, Mary C., Jagadeeswaran, Pudur
• Format: Article
• Language: English
• Subjects: Animals; Blood Platelets - metabolism; Breeding; Cyclic AMP; Danio rerio; DNA Mutational Analysis; Gene Expression; Genetics; haemostasis; Hematology; Injury analysis; Intravenous administration; Linkage analysis; Mutation; Occlusion; Peptide mapping; Phenotype; Platelets; Receptors, Lysophospholipid - genetics; Thrombocytes; Thromboembolism; Thrombosis; Zebrafish
• ISSN: 0007-1048; 1365-2141
• DOI: 10.1111/bjh.15046

Summary Haemostasis is a defence mechanism that has evolved to protect organisms from losing their circulating fluid. We have previously introduced zebrafish as a model to study the genetics of haemostasis to identify novel genes that play a role in haemostasis. Here, we identify a zebrafish mutant that showed prolonged time to occlusion (TTO) in the laser injury venous thrombosis assay. By linkage analysis and fine mapping, we found a mutation in the orphan G protein‐coupled receptor 34 like gene (gpr34l) causing a change of Val to Glu in the third external loop of Gpr34l. We have shown that injection of zebrafish gpr34l RNA rescues the prolonged TTO defect. The thrombocytes from the mutant showed elevated levels of cAMP that supports the defective thrombocyte function. We also have demonstrated that knockdown of this gene by intravenous Vivo‐Morpholino injections yielded a phenotype similar to the gpr34l mutation. These results suggest that the lack of functional Gpr34l leads to increased cAMP levels that result in defective thrombocyte aggregation.