Equine thrombospondin II and secreted protein acidic and cysteine-rich in a model of normal and pathological wound repair.
Abstract: Wound healing in horses is complicated, particularly when wounds are on the limb. The objectives of this study were to clone equine thrombospondin II (THBS2) and secreted protein acidic and cysteine-rich (SPARC) cDNAs and to compare the spatiotemporal expression of mRNAs and proteins during repair of body and limb wounds. These molecules were targeted in view of their potential biological contribution to angiogenesis, which is exacerbated during the repair of limb wounds in horses. Cloning was achieved by screening size-selected cDNA libraries previously derived from 7-day-old wounds. Expression was studied in unwounded skin and in samples from 1, 2, 3, 4, and 6 wk old wounds of the body and limb. Temporal gene expression was determined by semiquantitative RT-PCR, while protein expression was mapped immunohistochemically. The temporal pattern of expression for both genes was similar; wounding caused immediate upregulation of mRNA, which did not return to baseline by the end of the study, and overexpression was noted in body relative to limb wounds. Immunostaining for THBS2 and SPARC was induced by wounding, though no differences in stain location or intensity were detected between body and limb wounds. This study is the first to characterize equine cDNA for THBS2 and SPARC and to document mRNA expression over the different phases of repair. THBS2 and SPARC might modulate angiogenesis during wound healing in the horse, which could protect against the disproportionate fibroplasia commonly afflicting limb wounds and leading to the development of exuberant granulation tissue.
Publication Date: 2009-04-28 PubMed ID: 19401403DOI: 10.1152/physiolgenomics.90383.2008Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research paper explores how equine thrombospondin II (THBS2) and secreted protein acidic and cysteine-rich (SPARC) behave in horse wound healing, particularly in wounds located on the limb, which can lead to complications. The study involves cloning these proteins and investigating their expression during different stages of wound repair.
Objectives
- The research aimed at cloning the equine thrombospondin II (THBS2) and the secreted protein acidic and cysteine-rich (SPARC) cDNAs. These proteins were chosen due to their potential contribution to angiogenesis, a process that plays a significant role in healing limb wounds in horses.
- The study also sought to compare the spatiotemporal expression of these proteins’ mRNAs during the repair of body and limb wounds. This comparison was made by examining unwounded skin and wound samples that were 1, 2, 3, 4, and 6 weeks old.
Methods
- The cloning of THBS2 and SPARC was accomplished through screening size-selected cDNA libraries derived from 7-day-old wounds.
- To examine the temporal expression of these proteins, the research used semiquantitative RT-PCR to determine gene expression. Immunohistochemistry was used to map protein expression.
Results
- The results showed a similar temporal pattern of expression for both THBS2 and SPARC genes. There was an immediate upregulation of mRNA expression upon wounding which did not return to baseline levels by the end of the study.
- It was also observed that both proteins were overexpressed in body wounds compared to limb wounds.
- Immunostaining for THBS2 and SPARC was triggered by wounding, and no notable differences in staining location or intensity were observed between body and limb wounds.
Conclusions
- This study is the first to characterize equine cDNA for THBS2 and SPARC, as well as document mRNA expression during different repair phases.
- The results suggest that THBS2 and SPARC might modulate angiogenesis during wound healing in horses. This modulation could potentially protect against disproportionate fibroplasia, a condition that commonly affects limb wounds and leads to the formation of exuberant granulation tissue.
Cite This Article
APA
Miragliotta V, Raphaël K, Ipiña Z, Lussier JG, Theoret CL.
(2009).
Equine thrombospondin II and secreted protein acidic and cysteine-rich in a model of normal and pathological wound repair.
Physiol Genomics, 38(2), 149-157.
https://doi.org/10.1152/physiolgenomics.90383.2008 Publication
Researcher Affiliations
- Department of Veterinary Anatomy, Biochemistry and Physiology, University of Pisa, Pisa, Italy.
MeSH Terms
- Animals
- Cell Adhesion Molecules / genetics
- Cell Adhesion Molecules / metabolism
- Cloning, Molecular
- DNA Primers / genetics
- DNA, Complementary / genetics
- Horses
- Immunoblotting
- Immunohistochemistry
- Linear Models
- Neovascularization, Physiologic / genetics
- Neovascularization, Physiologic / physiology
- Osteonectin / genetics
- Osteonectin / metabolism
- RNA, Messenger / metabolism
- Reverse Transcriptase Polymerase Chain Reaction
- Thrombospondins / genetics
- Thrombospondins / metabolism
- Wound Healing / physiology
Citations
This article has been cited 3 times.- Baird A, Lindsay T, Everett A, Iyemere V, Paterson YZ, McClellan A, Henson FMD, Guest DJ. Osteoblast differentiation of equine induced pluripotent stem cells. Biol Open 2018 May 10;7(5).
- Gregori M, Miragliotta V, Leotta R, Cecchini S, Prearo M, Abramo F. Morphometric Evaluation of Interrenal Gland and Kidney Macrophages Aggregates in Normal Healthy Rainbow Trout (Oncorhynchus mykiss) and after Bacterial Challenge with Yersinia ruckeri. Vet Med Int 2014;2014:210625.
- Lee J, Bottje WG, Kong BW. Genome-wide host responses against infectious laryngotracheitis virus vaccine infection in chicken embryo lung cells. BMC Genomics 2012 Apr 24;13:143.
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