Detection of differentially regulated genes in ischaemic equine intestinal mucosa.
Abstract: Colic is a serious disease syndrome in horses. Much of the mortality is associated with ischaemic-injured intestine during strangulating obstruction, yet there is limited understanding of the associated molecular events. Identification of differentially expressed genes during ischaemic injury should expand our understanding of colic and may lead to novel targeted therapeutic approaches in the future. Objective: To isolate and identify differentially expressed genes in equine jejunum following a 2 h ischaemic event compared to normally perfused jejunum. Methods: Suppressive subtractive hybridisation was used to clone genes that are differentially expressed in equine jejunum injured by 2 h of complete ischaemia as compared to time-matched control jejunal tissues. Expression of selected clones was further evaluated by northern blot analysis. Results: Of the 384 clones selected, 157 were confirmed to possess cDNAs corresponding differentially expressed genes by dot blot analysis. Two genes, fatty acid binding protein 2 and calcium-activated chloride channel 4 were further confirmed to be differentially expressed by northern blot analysis. Conclusions: Suppressive subtractive hybridisation can be used to detect changes in expression of a broad array of genes, as confirmed by northern blot analysis of selected genes. Conclusions: These initial results have identified a pool of equine intestinal epithelial genes that are differentially expressed following a 2 h ischaemic event. In particular, genes indicative of deranged metabolic activity and those potentially involved in early repair events were identified and may ultimately provide clues as to the nature of epithelial ischaemic injury in horses.
Publication Date: 2005-07-21 PubMed ID: 16028620DOI: 10.2746/0425164054529382Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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The research article explores the identification of genes that are differently expressed in horse’s intestine due to ischaemic injury. This understanding is crucial to expanding our knowledge about colic, a serious condition in horses, and potentially developing new targeted treatments in the future.
Research Objective
- The main goal of this study was to pinpoint and identify genes whose expression varies in the jejunum of horses after a two-hour ischaemic incident as compared to a normally functioning jejunum.
Methods Employed
- Suppressive subtractive hybridisation, a technique used to identify unique sequences of nucleic acids, was used by researchers to clone genes with differential expression in the jejunum that had undergone 2 hours of complete ischaemia – compared to control tissues which were time-matched.
- To validate this further, selected clones’ expression was studied via northern blot analysis, a documented technique for detecting specific RNA sequences within a complex mixture.
Research Findings
- Out of the total 384 cloned genes, 157 were verified to possess cDNAs corresponding differentially expressed genes by dot blot analysis – a technique used to detect, analyse and identify proteins present in a sample.
- Further verification by northern blot analysis detected that two particular genes, fatty acid binding protein 2 and calcium-activated chloride channel 4, were indeed differentially expressed.
Conclusions
- Suppressive subtractive hybridisation was deemed to be effective in discerning changes in the expression of a wide variety of genes, as corroborated by northern blot analysis.
- The initial findings revealed a cluster of equine intestinal epithelial genes that expressed differently after a 2-hour ischaemic incident. Among these, genes that reflected disordered metabolic activity and those perhaps involved in early repair events were identified. This could provide clues about the nature of epithelial ischaemic injury in horses.
Cite This Article
APA
Tschetter JR, Blikslager AT, Little D, Howard RD, Woody SL, Beex LM, Crisman MV.
(2005).
Detection of differentially regulated genes in ischaemic equine intestinal mucosa.
Equine Vet J, 37(4), 319-324.
https://doi.org/10.2746/0425164054529382 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, Virginia Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061, USA.
MeSH Terms
- Animals
- Base Sequence
- Blotting, Northern / veterinary
- Cloning, Molecular
- Colic / etiology
- Colic / veterinary
- Gene Expression Profiling
- Gene Expression Regulation
- Gene Library
- Horse Diseases / genetics
- Horses
- Immunoblotting / veterinary
- Intestinal Mucosa / blood supply
- Intestinal Mucosa / metabolism
- Ischemia / genetics
- Ischemia / veterinary
- Jejunum / blood supply
- Male
- RNA / metabolism
Citations
This article has been cited 1 times.- Marañón G, Manley W, Cayado P, García C, de la Muela MS, Vara E. Alterations in the glutathione metabolism could be implicated in the ischemia-induced small intestinal cell damage in horses. BMC Vet Res 2009 Mar 18;5:10.
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