Exploring erythropoiesis of common carp (Cyprinus carpio) using an in vitro colony assay in the presence of recombinant carp kit ligand A and erythropoietin. Issue 1 (November 2015)

• Record Type: Journal Article
• Authors: Katakura, Fumihiko; Yabu, Takeshi; Yamaguchi, Takuya; Miyamae, Jiro; Shirinashihama, Yuki; Nakanishi, Teruyuki; Moritomo, Tadaaki
• Is Part Of: Developmental and comparative immunology. Volume 53:Issue 1(2015)
• Page Start: 13
• Page End: 22
• Publication Date: 2015-11
• Subjects: Erythropoiesis -- Colony assay -- Teleost -- Erythropoietin -- Kit ligand A; Immunology -- Periodicals; Developmental immunology -- Periodicals; 616.079
• DOI: 10.1016/j.dci.2015.06.006
• Languages: English
• ISSNs: 0145-305X

Abstract

Abstract: The use of in vitro colony assays in mammals has contributed to identification of erythroid progenitor cells such as burst-forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E) progenitors, and serves to examine functions of erythropoietic growth factors like Erythropoietin (Epo) and Kit ligand. Here, we established an in vitro colony-forming assay capable of investigating erythropoiesis in carp ( Cyprinus carpio ), cloned and functionally characterized recombinant homologous molecules Epo and Kit ligand A (Kitla), and identified three distinct erythroid progenitor cells in carp. Recombinant carp Epo induced the formation of CFU-E-like and BFU-E-like erythroid colonies, expressing erythroid marker genes, β-globin, epor and gata1 . Recombinant carp Kitla alone induced limited colony formation, whereas a combination of Kitla and Epo dramatically enhanced erythroid colony formation and colony cell growth, as well as stimulated the formation of thrombocytic/erythroid colonies expressing not only erythroid markers but also thrombocytic markers, cd41 and c-mpl . Utilizing this colony assay to examine the distribution of distinct erythroid progenitor cells in carp, we demonstrated that carp head and trunk kidney play a primary role in erythropoiesis, while the spleen plays a secondary. Furthermore, we showed that presumably bi-potent thrombocytic/erythroid progenitor cells localize principally in the trunk kidney. Our results indicate that teleost fish possess mechanisms of Epo- and Kitla-dependent erythropoiesis similar to those in other vertebrates, and also help to demonstrate the diversity of erythropoietic sites among vertebrates. Highlights: Recombinant carp erythropoietin induced the formation of CFU-E-like and BFU-E-like erythroid colonies. Kit ligand A in cooperation with erythropoietin synergistically promote erythroid and thrombocytic/erythroid colony formation. Carp head and trunk kidney play a primary role in erythropoiesis, while the spleen plays a secondary.