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Aging Markers in Equine Red Blood Cells

Detection of hematopoietic activity in horses is a challenge due to the lack of cells carrying reticulocyte markers such as RNA remnants or CD71 in the circulation. In this study, we fractionated equine red cells according to their density and analyzed the cells forming low (L), medium (M), and high...

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Autores principales: Kämpf, Sandra, Seiler, Elena, Bujok, Jolanta, Hofmann-Lehmann, Regina, Riond, Barbara, Makhro, Asya, Bogdanova, Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6650539/
https://www.ncbi.nlm.nih.gov/pubmed/31379601
http://dx.doi.org/10.3389/fphys.2019.00893
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author Kämpf, Sandra
Seiler, Elena
Bujok, Jolanta
Hofmann-Lehmann, Regina
Riond, Barbara
Makhro, Asya
Bogdanova, Anna
author_facet Kämpf, Sandra
Seiler, Elena
Bujok, Jolanta
Hofmann-Lehmann, Regina
Riond, Barbara
Makhro, Asya
Bogdanova, Anna
author_sort Kämpf, Sandra
collection PubMed
description Detection of hematopoietic activity in horses is a challenge due to the lack of cells carrying reticulocyte markers such as RNA remnants or CD71 in the circulation. In this study, we fractionated equine red cells according to their density and analyzed the cells forming low (L), medium (M), and high (H) density fractions for markers of aging such as membrane loss, oxidation, and alterations in the intracellular free Ca(2+) levels. Cells forming L and M fraction were highly heterogeneous in projected areas and shapes, and had higher propensity to swell in response to hypo-osmotic challenge than the cells from the H fraction. The densest cells were deprived of band 3 protein compared to the cells within L or M fraction. Furthermore, the equine red cells from the H fraction were hyper-oxidized compared to the cells within M and L fractions as follows from an increase in autofluorescence characteristic for oxidized damaged hemoglobin and from thiol oxidation as detected using monobromobimane. The lightest cells showed lower free thiol content compared to the red blood cells from the M fraction, but did not contain oxidized hemoglobin. Finally, the majority of red blood cells forming L, M, and H fraction prominently differed from each other in intracellular free Ca(2+) levels and its distribution within the cells. Based on the obtained findings, we suggest that intraerythrocytic Ca(2+) levels and its subcellular distribution, eosin-5-maleimide binding test for band 3 abundance, and autofluorescence of cells along with the changes in red blood cell indices, distribution width and creatine levels may become potential markers of regenerative erythropoiesis in horses. Validation of the power of these potential markers of red cell aging is pending.
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spelling pubmed-66505392019-08-02 Aging Markers in Equine Red Blood Cells Kämpf, Sandra Seiler, Elena Bujok, Jolanta Hofmann-Lehmann, Regina Riond, Barbara Makhro, Asya Bogdanova, Anna Front Physiol Physiology Detection of hematopoietic activity in horses is a challenge due to the lack of cells carrying reticulocyte markers such as RNA remnants or CD71 in the circulation. In this study, we fractionated equine red cells according to their density and analyzed the cells forming low (L), medium (M), and high (H) density fractions for markers of aging such as membrane loss, oxidation, and alterations in the intracellular free Ca(2+) levels. Cells forming L and M fraction were highly heterogeneous in projected areas and shapes, and had higher propensity to swell in response to hypo-osmotic challenge than the cells from the H fraction. The densest cells were deprived of band 3 protein compared to the cells within L or M fraction. Furthermore, the equine red cells from the H fraction were hyper-oxidized compared to the cells within M and L fractions as follows from an increase in autofluorescence characteristic for oxidized damaged hemoglobin and from thiol oxidation as detected using monobromobimane. The lightest cells showed lower free thiol content compared to the red blood cells from the M fraction, but did not contain oxidized hemoglobin. Finally, the majority of red blood cells forming L, M, and H fraction prominently differed from each other in intracellular free Ca(2+) levels and its distribution within the cells. Based on the obtained findings, we suggest that intraerythrocytic Ca(2+) levels and its subcellular distribution, eosin-5-maleimide binding test for band 3 abundance, and autofluorescence of cells along with the changes in red blood cell indices, distribution width and creatine levels may become potential markers of regenerative erythropoiesis in horses. Validation of the power of these potential markers of red cell aging is pending. Frontiers Media S.A. 2019-07-17 /pmc/articles/PMC6650539/ /pubmed/31379601 http://dx.doi.org/10.3389/fphys.2019.00893 Text en Copyright © 2019 Kämpf, Seiler, Bujok, Hofmann-Lehmann, Riond, Makhro and Bogdanova. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Kämpf, Sandra
Seiler, Elena
Bujok, Jolanta
Hofmann-Lehmann, Regina
Riond, Barbara
Makhro, Asya
Bogdanova, Anna
Aging Markers in Equine Red Blood Cells
title Aging Markers in Equine Red Blood Cells
title_full Aging Markers in Equine Red Blood Cells
title_fullStr Aging Markers in Equine Red Blood Cells
title_full_unstemmed Aging Markers in Equine Red Blood Cells
title_short Aging Markers in Equine Red Blood Cells
title_sort aging markers in equine red blood cells
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6650539/
https://www.ncbi.nlm.nih.gov/pubmed/31379601
http://dx.doi.org/10.3389/fphys.2019.00893
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