title: Exploring erythropoiesis of common carp (Cyprinus carpio) using an in vitro colony assay in the presence of recombinant carp kit ligand A and erythropoietin.
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.
authors.affiliation: Laboratory of Comparative Immunology, Department of Veterinary Medicine, Nihon University, Kameino 1866, Fujisawa, Kanagawa 252-0880, Japan. Electronic address: katakura.fumihiko@nihon-u.ac.jp.
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Katakura
authors.affiliation: Laboratory of Fish Pathology, Department of Veterinary Medicine, Nihon University, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yabu
authors.affiliation: Laboratory of Fish Pathology, Department of Veterinary Medicine, Nihon University, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Yamaguchi
authors.affiliation: Laboratory of Comparative Immunology, Department of Veterinary Medicine, Nihon University, Kameino 1866, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Jiro
authors.initials: J
authors.lastname: Miyamae
authors.affiliation: Laboratory of Fish Pathology, Department of Veterinary Medicine, Nihon University, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Shirinashihama
authors.affiliation: Laboratory of Fish Pathology, Department of Veterinary Medicine, Nihon University, Japan.
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nakanishi
authors.affiliation: Laboratory of Comparative Immunology, Department of Veterinary Medicine, Nihon University, Kameino 1866, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Tadaaki
authors.initials: T
authors.lastname: Moritomo
mesh_terms: D000818:Animals; D002347:Carps; D004920:Erythropoiesis / Q000502:physiology*; D004921:Erythropoietin / Q000235:genetics*; D050982:GATA1 Transcription Factor / Q000096:biosynthesis; D007668:Kidney / Q000378:metabolism; D040201:Platelet Membrane Glycoprotein IIb / Q000096:biosynthesis; D017467:Receptors, Erythropoietin / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000235:genetics; D013154:Spleen / Q000378:metabolism; D019089:Stem Cell Factor / Q000235:genetics*; D013234:Stem Cells / Q000166:cytology*; D013926:Thrombopoietin / Q000096:biosynthesis; D055544:beta-Globins / Q000096:biosynthesis
chemical_list: D050982:GATA1 Transcription Factor; D040201:Platelet Membrane Glycoprotein IIb; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D055544:beta-Globins; D004921:Erythropoietin; D013926:Thrombopoietin
keywords: Colony assay; Erythropoiesis; Erythropoietin; Kit ligand A; Teleost