PUBLICATION
Unconventional interaction of the zebrafish thrombopoietin receptor with Mapk14a determines thrombocyte development and function
- Authors
- Thakkar, K.J., Naseer, S., Chowdhury, S., Mondal, S., Shrivastva, S., Ribeyron, L., Menon, M.B., Kapuria, V., Minocha, S., Roy, A.
- ID
- ZDB-PUB-250317-11
- Date
- 2025
- Source
- Journal of thrombosis and haemostasis : JTH : (Journal)
- Registered Authors
- Keywords
- Mapk14a, Zebrafish, signalling, thrombocytes, thrombopoietin receptor
- MeSH Terms
- none
- PubMed
- 40089183 Full text @ J. Thromb. Haemost.
Citation
Thakkar, K.J., Naseer, S., Chowdhury, S., Mondal, S., Shrivastva, S., Ribeyron, L., Menon, M.B., Kapuria, V., Minocha, S., Roy, A. (2025) Unconventional interaction of the zebrafish thrombopoietin receptor with Mapk14a determines thrombocyte development and function. Journal of thrombosis and haemostasis : JTH. :. Epub ahead of print.
Abstract
Background Thrombopoietin receptor signalling permits the thrombocyte lineage development in human, mice as well as zebrafish. While the sequence, structure, signalling, and function of TpoR is well-conserved in humans and mice, the zebrafish thrombopoietin receptor (zTpoR) remains less understood.
Objectives In this article, we identified the interactome of the cytoplasmic domain of zTpoR.
Methods Using recombinant zTpoR cytoplasmic domain, we designed a pull down assay combined with mass spectrometry to identify possible interactors of the cytoplasmic domain. We further used biochemistry, flow cytometry and zebrafish bleeding time assay to demonstrate the effects of these interactions.
Results We observed a specific interaction of the zTpoR with Mapk14a, a homolog of the human and mouse p38 MAPKα. Interestingly, this interaction was not observed with human TpoR, highlighting a unique aspect of zebrafish biology. To understand the physiological consequence of zTpoR - Mapk14a interaction, we used inhibitors of Jak (ruxolitinib) and Mapk14a (BIRB 796) in zebrafish. Both ruxolitinib and BIRB 796 reduced the thrombocyte, myeloid and progenitor cell count while increasing the bleeding time. Moreover, combinatorial treatment with BIRB 796 and ruxolitinib showed an additive effect on bleeding time without any further decrease in the myeloid and progenitor cell counts.
Conclusion We thus identified and characterized a specific interaction of zTpoR with Mapk14a that regulates zebrafish thrombopoiesis in both embryonic and adult stages.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping