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Cloning and tissue expression of the equine transferrin receptor.
Abstract: Characterization of anemia in horses presents a challenge, as they do not release reticulocytes into peripheral blood. Transferrin receptor (TfR) expression is highest on erythroid cells in people and rats, and measurement of a soluble serum form (sTfR) is used to quantify erythropoiesis in these species. We hypothesized that equine TfR (eTfR) expression is similar in quantity and distribution to that in these other species and thus has potential for characterization of the regenerative response in anemic horses. Objective: This study was conducted to clone and sequence the eTfR gene and measure expression levels using quantitative real-time PCR and immunohistochemical (IHC) staining. Methods: Total RNA from equine bone marrow was used to produce cDNA. The eTfR gene was amplified using pooled gene-specific primers, and PCR products were sequenced. Rapid amplification of cDNA ends was used to obtain the first 22 nucleotides of the coding sequence. Quantitative PCR was performed using eTfR gene-specific primers, and IHC staining was used to localize TfR protein expression. Results: The deduced amino acid (aa) sequence (767 aa) of the eTfR was 75-83% identical with sequences of the receptor in several other mammals. As in people and rats, eTfR mRNA expression was highest in the bone marrow, and distribution in other tissues was also similar. Conclusions: The eTfR gene is similar to that of other mammals in structure and expression levels. We hypothesize that it is also similar in function and that, following development of an immunoassay, determining sTfR concentrations will be useful for identifying the regenerative response in anemic horses.
©2010 American Society for Veterinary Clinical Pathology.
Publication Date: 2010-11-11 PubMed ID: 21070303DOI: 10.1111/j.1939-165X.2010.00265.xGoogle Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research focuses on the study of the equine transferrin receptor (eTfR) which exhibits similar properties to its counterparts in other species. Information gleaned from the study could potentially help understand and measure the regenerative response in horses suffering from anemia.
Objective and Methods of Research
The primary objective of the research was to clone and sequence the eTfR gene and measure expression levels using two methods:
- Quantitative real-time PCR (polymerase chain reaction) which is an advanced technique used to copy and amplify minute quantities of DNA.
- Immunohistochemical (IHC) staining which is a method to detect antigens (proteins) in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues.
To proceed with their experiment, the researchers used total RNA from equine bone marrow to produce cDNA (complementary DNA). The eTfR gene was then amplified using gene-specific primers, and PCR products were sequenced. Additionally, a technique called “rapid amplification of cDNA ends” was used to derive the first 22 nucleotides of the coding sequence.
Results of the Study
The researchers noticed that the deduced amino acid sequence of the eTfR was 75-83% identical with sequences of the receptor in several other mammals. The eTfR mRNA expression was highest in the bone marrow in horses, akin to people and rats, and distribution in other tissues was also similarity observed.
Conclusion and Implication
Based on the results, the study concluded that the eTfR gene in horses is similar to that of other mammals regarding its structure and expression levels. Given these similarities, it was inferred that the eTfR gene would exhibit similar functionalities as well. This conclusion opens the door to the development of an immunoassay – a biochemical test that measures the presence or concentration of a substance in solutions that frequently contain a complex mixture of substances. Through such an immunoassay, determining sTfR (soluble TfR) concentrations might help identify the regenerative response in anemic horses, meaning this could serve as a novel method for diagnosing and assessing treatment for equine anemia.
Cite This Article
APA
Webb TL, Burnett RC, Avery AC, Olver CS.
(2010).
Cloning and tissue expression of the equine transferrin receptor.
Vet Clin Pathol, 39(4), 424-432.
https://doi.org/10.1111/j.1939-165X.2010.00265.x Publication
Researcher Affiliations
- Department of Microbiology, Colorado State University, Fort Collins, CO, USA.
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Cloning, Molecular
- Genes / genetics
- Horses / genetics
- Molecular Sequence Data
- RNA, Messenger / genetics
- Receptors, Transferrin / biosynthesis
- Receptors, Transferrin / genetics
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Sequence Alignment
- Tissue Distribution
Citations
This article has been cited 1 times.- Kaelber JT, Demogines A, Harbison CE, Allison AB, Goodman LB, Ortega AN, Sawyer SL, Parrish CR. Evolutionary reconstructions of the transferrin receptor of Caniforms supports canine parvovirus being a re-emerged and not a novel pathogen in dogs. PLoS Pathog 2012;8(5):e1002666.
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