Transferrin receptor expression in serum exosomes as a marker of regenerative anaemia in the horse.
Abstract: Evaluation of erythrocyte regeneration in horses is challenging, as they do not release reticulocytes into the peripheral blood. This study investigated transferrin receptor 1 (TfR1) expression in exosomes as a noninvasive method of characterising the regenerative response in anaemic horses. Objective: To quantify TfR1 in ultraprecipitate of serum in horses before and after phlebotomy-induced anaemia, and to identify exosomes as the source of TfR1. The hypothesis was that serum exosomal TfR1 expression would increase during a regenerative response. Methods: Experimental model of anaemia. Methods: Six horses were phlebotomised to achieve a 25% decrease in packed cell volume. Transferrin receptor 1 quantity in exosomes was determined by western blot and relative densitometry before and after phlebotomy. The size and density of the TfR1-associated particles were confirmed by transmission electron microscopy and density gradient centrifugation, respectively. Results: Regenerative anaemia was confirmed by decreased packed cell volumes and decreased myeloid:erythroid ratios in the bone marrow. In all 6 horses, TfR1 expression increased between Days 7 and 10. Mean TfR1 levels peaked on Day 10 and at 3-fold higher than levels on Day 0. Appropriately sized particles were evident on transmission electron microscopy and sucrose density gradient fractions expected to contain exosomes also contained TfR1. Conclusions: These data indicate that TfR1 expression in serum exosomes may provide a marker for regeneration in anaemic horses.
© 2014 EVJ Ltd.
Publication Date: 2014-03-13 PubMed ID: 24708277DOI: 10.1111/evj.12235Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the use of transferrin receptor 1 (TfR1) expression in serum exosomes as a potential marker of red blood cell regeneration (erythrocyte regeneration) in anaemic horses. It demonstrates that TfR1 levels in serum exosomes might be useful for determining the regenerative response in horses when they suffer from anaemia.
Objective and Hypothesis
- The primary objective of this study was to measure the quantity of transferrin receptor 1 (TfR1) in the ultraprecipitate of serum in horses before and after they were made anaemic through phlebotomy (blood withdrawal)
- The researchers hypothesized that the expression of TfR1 in serum exosomes would increase during a regenerative response to anaemia
Research Strategy
- This study used an experimental model of anaemia in which six horses were intentionally made anaemic to effect a 25% decrease in their packed cell volume (a measure of the volume of red blood cells in the blood).
- The amount of TfR1 in exosomes was determined by western blot and relative densitometry before and after the horses were made anaemic.
- The size and density of the particles associated with TfR1 were confirmed using transmission electron microscopy and density gradient centrifugation, respectively.
Results
- After inducing anaemia, a decrease was seen in packed cell volumes and in myeloid:erythroid ratios in the bone marrow, which confirms that regenerative anaemia had been induced.
- Following anaemia induction, TfR1 expression increased between Days 7 and 10 in all horses.
- Near tripling of the mean TfR1 levels was observed on Day 10 compared to levels on Day 0.
- Particles of the appropriate size for exosomes as well the presence of TfR1 in sucrose density gradient fractions expected to contain exosomes were evident when examined using electron microscopy and centrifugation.
Conclusion
- The increase in TfR1 expression observed in serum exosomes during the regenerative response to anaemia supports the use of this marker to assess regenerative anaemia in horses.
Cite This Article
APA
Rout ED, Webb TL, Laurence HM, Long L, Olver CS.
(2014).
Transferrin receptor expression in serum exosomes as a marker of regenerative anaemia in the horse.
Equine Vet J, 47(1), 101-106.
https://doi.org/10.1111/evj.12235 Publication
Researcher Affiliations
- Fort Collins Veterinary Emergency and Rehabilitation Hospital, Fort Collins, Colorado, USA.
MeSH Terms
- Anemia / blood
- Anemia / metabolism
- Anemia / veterinary
- Animals
- Biomarkers
- Exosomes / metabolism
- Female
- Gene Expression Regulation / physiology
- Horse Diseases / blood
- Horse Diseases / metabolism
- Horses
- Male
- Receptors, Transferrin / blood
- Receptors, Transferrin / genetics
- Receptors, Transferrin / metabolism
Citations
This article has been cited 9 times.- Moccia V, Sammarco A, Cavicchioli L, Castagnaro M, Bongiovanni L, Zappulli V. Extracellular Vesicles in Veterinary Medicine. Animals (Basel) 2022 Oct 10;12(19).
- Tang X, Chen F, Xie LC, Liu SX, Mai HR. Targeting metabolism: A potential strategy for hematological cancer therapy. World J Clin Cases 2022 Apr 6;10(10):2990-3004.
- Bongiovanni L, Andriessen A, Wauben MHM, Nolte-'t Hoen ENM, de Bruin A. Extracellular Vesicles: Novel Opportunities to Understand and Detect Neoplastic Diseases. Vet Pathol 2021 May;58(3):453-471.
- Kämpf S, Seiler E, Bujok J, Hofmann-Lehmann R, Riond B, Makhro A, Bogdanova A. Aging Markers in Equine Red Blood Cells. Front Physiol 2019;10:893.
- Burkova EE, Grigor'eva AE, Bulgakov DV, Dmitrenok PS, Vlassov VV, Ryabchikova EI, Sedykh SE, Nevinsky GA. Extra Purified Exosomes from Human Placenta Contain An Unpredictable Small Number of Different Major Proteins. Int J Mol Sci 2019 May 16;20(10).
- Sedykh SE, Purvinish LV, Monogarov AS, Burkova EE, Grigor'eva AE, Bulgakov DV, Dmitrenok PS, Vlassov VV, Ryabchikova EI, Nevinsky GA. Purified horse milk exosomes contain an unpredictable small number of major proteins. Biochim Open 2017 Jun;4:61-72.
- Carossino M, Dini P, Kalbfleisch TS, Loynachan AT, Canisso IF, Shuck KM, Timoney PJ, Cook RF, Balasuriya UBR. Downregulation of MicroRNA eca-mir-128 in Seminal Exosomes and Enhanced Expression of CXCL16 in the Stallion Reproductive Tract Are Associated with Long-Term Persistence of Equine Arteritis Virus. J Virol 2018 May 1;92(9).
- Nagarajah S. Exosome Secretion - More Than Simple Waste Disposal? Implications for Physiology, Diagnostics and Therapeutics. J Circ Biomark 2016 Jan-Dec;5:7.
- González E, Falcón-Pérez JM. Expanding Horizons: Next-Generation and Interdisciplinary Advances in the Applications of Extracellular Vesicles. J Extracell Biol 2025 Dec;4(12):e70101.
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