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Frontiers in physiology2019; 10; 893; doi: 10.3389/fphys.2019.00893

Aging Markers in Equine Red Blood Cells.

Abstract: 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 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 levels and its distribution within the cells. Based on the obtained findings, we suggest that intraerythrocytic Ca 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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Publication Date: 2019-07-17 PubMed ID: 31379601PubMed Central: PMC6650539DOI: 10.3389/fphys.2019.00893Google Scholar: Lookup
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Summary

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This research investigates potential markers of aging in horse red blood cells by fractionating them according to density and assessing for changes in markers such as membrane loss, oxidation, and alterations in intracellular free Calcium levels. The study proposes that monitoring of these markers, along with changes in red blood cell indices, distribution width and creatine levels, may be useful in detecting regenerative erythropoiesis in horses.

About the Study

The primary objective of this research was to identify potential markers of red blood cell (RBC) aging in horses, which may eventually be used to detect regenerative erythropoiesis.

  • The authors compared red blood cells fractioned into three different density groups – low (L), medium (M), and high (H), and analyzed these fractions for various aging markers.
  • Erythropoiesis is the process of producing new red blood cells. Detection of this activity, particularly regenerative erythropoiesis (which kicks in when there is an increased demand for new red blood cells such as in the case of blood loss or anemia), has been challenging in horses due to the lack of traditional hematopoietic markers in their circulation.

The Findings

  • Cells in the L and M fractions were heterogeneous and prone to swell in response to a hypo-osmotic challenge, suggesting these cells may be older and more susceptible to osmotic stress.
  • The H fraction cells had less of a protein called ‘band 3’, which is important to red cell function, compared to L and M fractions. This suggests that these denser cells may be aged or damaged in some way.
  • Cells in the H fraction were more oxidized, which is a common feature of aging or damaged cells. This was deduced by observing an increase in autofluorescence and thiol oxidation, which are markers for oxidized or damaged hemoglobin.
  • The L fraction cells showed decreased free thiol content compared to M fraction cells, which is typically indicative of changes in the cellular redox state associated with aging. However, these lightest cells were absent of oxidized hemoglobin, highlighting their complex and diverse nature.
  • The level of intracellular free Calcium varied prominently across all fractions, along with its distribution within the cells, suggesting that calcium levels may be a potential aging indicator.

Potential Markers and Further Research

Based on these observations, the authors proposed several potential markers for red cell aging and regenerative erythropoiesis in horses, including:

  • Intraerythrocytic Calcium levels and its subcellular distribution
  • Eosin-5-maleimide binding test for band 3 abundance
  • Cellular autofluorescence
  • Alterations in red blood cell indices, distribution width, and creatine levels

The study acknowledges the need for further investigations and validation to confirm the utility of these proposed markers for red blood cell aging in horses.

Cite This Article

APA
Kämpf S, Seiler E, Bujok J, Hofmann-Lehmann R, Riond B, Makhro A, Bogdanova A. (2019). Aging Markers in Equine Red Blood Cells. Front Physiol, 10, 893. https://doi.org/10.3389/fphys.2019.00893

Publication

Frontiers in physiology
ISSN: 1664-042X
NlmUniqueID: 101549006
Country: Switzerland
Language: English
Volume: 10
Pages: 893
PII: 893

Researcher Affiliations

Kämpf, Sandra
  • Red Blood Cell Research Group, Vetsuisse Faculty, Institute of Veterinary Physiology, University of Zurich, Zürich, Switzerland.
  • Vetsuisse Faculty, University of Bern, Bern, Switzerland.
Seiler, Elena
  • Red Blood Cell Research Group, Vetsuisse Faculty, Institute of Veterinary Physiology, University of Zurich, Zürich, Switzerland.
Bujok, Jolanta
  • Red Blood Cell Research Group, Vetsuisse Faculty, Institute of Veterinary Physiology, University of Zurich, Zürich, Switzerland.
  • Institute of Animal Physiology, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
Hofmann-Lehmann, Regina
  • Clinical Laboratory, Vetsuisse Faculty, University of Zurich, Zürich, Switzerland.
Riond, Barbara
  • Clinical Laboratory, Vetsuisse Faculty, University of Zurich, Zürich, Switzerland.
Makhro, Asya
  • Red Blood Cell Research Group, Vetsuisse Faculty, Institute of Veterinary Physiology, University of Zurich, Zürich, Switzerland.
Bogdanova, Anna
  • Red Blood Cell Research Group, Vetsuisse Faculty, Institute of Veterinary Physiology, University of Zurich, Zürich, Switzerland.
  • The Zurich Center for Integrative Human Physiology (ZIHP), Zürich, Switzerland.

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Citations

This article has been cited 5 times.
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