Protein kinase C alpha and beta are positive regulators of thrombus formation in vivo in a zebrafish (Danio rerio) model of thrombosis - PubMed (original) (raw)

Protein kinase C alpha and beta are positive regulators of thrombus formation in vivo in a zebrafish (Danio rerio) model of thrombosis

C M Williams et al. J Thromb Haemost. 2011 Dec.

Free article

Abstract

Background: The zebrafish (Danio rerio) is becoming an attractive model organism for the assessment of gene function in thrombosis in vivo. Zebrafish, as a thrombosis model, have several advantages, with the capacity to follow thrombus formation at high resolution in real time using intravital microscopy, without the need for complex surgical techniques, and the capability to rapidly knockdown gene expression using morpholino antisense approaches.

Objectives: We have recently shown, in mouse models, that protein kinase C alpha (PKCα) plays a critical role in regulating thrombus formation in vivo. PKC beta (β) plays a non-redundant role also in platelet function in vitro, but the function of this gene had not yet been assessed in vivo.

Methods: In the present study, we analyzed the function of both PKCα and PKCβ in the zebrafish model in vivo, by live imaging using a laser-induced injury of the main caudal artery in 3-day-old larvae.

Results: We showed that D. rerio express orthologs of both the PKCα and PKCβ genes, with high sequence identity. Translation blocking and splice-blocking morpholinos effectively and specifically knockdown expression of these genes and knockdown with either morpholino leads to attenuated thrombus formation, as assessed by several quantitative parameters including time to initial adhesion and peak thrombus surface area.

Conclusions: Our data indicate that these two highly related genes play non-redundant roles in regulating thrombosis, an observation that supports our previous in vitro murine data, and suggests unique roles, and possibly unique regulation, for PKCα and PKCβ in controlling platelet function in vivo.

PubMed Disclaimer

Cited by

Functional Genomics for the Identification of Modulators of Platelet-Dependent Thrombus Formation.
Vermeersch E, Nuyttens BP, Tersteeg C, Broos K, De Meyer SF, Vanhoorelbeke K, Deckmyn H. Vermeersch E, et al. TH Open. 2018 Sep 10;2(3):e272-e279. doi: 10.1055/s-0038-1670630. eCollection 2018 Jul. TH Open. 2018. PMID: 31249951 Free PMC article. Review.
Antithrombotic effect and action mechanism of Salvia miltiorrhiza and Panax notoginseng herbal pair on the zebrafish.
Yin SJ, Luo YQ, Zhao CP, Chen H, Zhong ZF, Wang S, Wang YT, Yang FQ. Yin SJ, et al. Chin Med. 2020 Apr 16;15:35. doi: 10.1186/s13020-020-00316-y. eCollection 2020. Chin Med. 2020. PMID: 32322295 Free PMC article.
Identification of the Antithrombotic Mechanism of Leonurine in Adrenalin Hydrochloride-Induced Thrombosis in Zebrafish via Regulating Oxidative Stress and Coagulation Cascade.
Liao L, Zhou M, Wang J, Xue X, Deng Y, Zhao X, Peng C, Li Y. Liao L, et al. Front Pharmacol. 2021 Nov 4;12:742954. doi: 10.3389/fphar.2021.742954. eCollection 2021. Front Pharmacol. 2021. PMID: 34803688 Free PMC article.
Cytohesin-2 phosphorylation by protein kinase C relieves the constitutive suppression of platelet dense granule secretion by ADP-ribosylation factor 6.
van den Bosch MT, Poole AW, Hers I. van den Bosch MT, et al. J Thromb Haemost. 2014 May;12(5):726-35. doi: 10.1111/jth.12542. J Thromb Haemost. 2014. PMID: 24581425 Free PMC article.
PKC-epsilon deficiency alters progenitor cell populations in favor of megakaryopoiesis.
Kostyak JC, Liverani E, Kunapuli SP. Kostyak JC, et al. PLoS One. 2017 Aug 4;12(8):e0182867. doi: 10.1371/journal.pone.0182867. eCollection 2017. PLoS One. 2017. PMID: 28783756 Free PMC article.

Protein kinase C alpha and beta are positive regulators of thrombus formation in vivo in a zebrafish (Danio rerio) model of thrombosis - PubMed (original) (raw)