Isolation and evaluation of erythroid progenitors in the livers of larval, froglet, and adult Xenopus tropicalis

Xenopus liver maintains erythropoietic activity from the larval to the adult stage. During metamorphosis, thyroid hormone mediates apoptosis of larval-type erythroid progenitors and proliferation of adult-type erythroid progenitors, and a globin switch occurs during this time. In addition, the whole...

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Bibliographic Details
Published in:Biology open 2023-08, Vol.12 (8)
Main Authors: Omata, Kazuki, Nomura, Ikki, Hirata, Akito, Yonezuka, Yuka, Muto, Hiroshi, Kuriki, Ryo, Jimbo, Kirin, Ogasa, Koujin, Kato, Takashi
Format: Article
Language:English
Subjects:
acridine orange
Aging
Animals
Cell Differentiation
Cell Separation
Erythroid Cells - metabolism
erythroid progenitor
Erythropoietin - metabolism
erythropoietin receptor
flow cytometry
HEK293 Cells
Humans
Larva - metabolism
Liver - metabolism
Receptors, Erythropoietin - metabolism
Xenopus
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Summary:Xenopus liver maintains erythropoietic activity from the larval to the adult stage. During metamorphosis, thyroid hormone mediates apoptosis of larval-type erythroid progenitors and proliferation of adult-type erythroid progenitors, and a globin switch occurs during this time. In addition, the whole-body mass and the liver also change; however, whether there is a change in the absolute number of erythroid progenitors is unclear. To isolate and evaluate erythroid progenitors in the Xenopus liver, we developed monoclonal ER9 antibodies against the erythropoietin receptor (EPOR) of Xenopus. ER9 recognized erythrocytes, but not white blood cells or thrombocytes. The specificity of ER9 for EPOR manifested as its inhibitory effect on the proliferation of a Xenopus EPOR-expressing cell line. Furthermore, ER9 recognition was consistent with epor gene expression. ER9 staining with Acridine orange (AO) allowed erythrocyte fractionation through fluorescence-activated cell sorting. The ER9+ and AO-red (AOr)high fractions were highly enriched in erythroid progenitors and primarily localized to the liver. The method developed using ER9 and AO was also applied to larvae and froglets with different progenitor populations from adult frogs. The liver to body weight and the number of ER9+ AOrhigh cells per unit body weight were significantly higher in adults than in larvae and froglets, and the number of ER9+ AOrhigh cells per unit liver weight was the highest in froglets. Collectively, our results show increased erythropoiesis in the froglet liver and demonstrate growth-dependent changes in erythropoiesis patterns in specific organs of Xenopus.
ISSN:2046-6390
2046-6390
DOI:10.1242/bio.059862