Detection of differentially regulated genes in ischaemic equine intestinal mucosa.
BACKGROUND
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.
Tschetter
J R
JR
Department of Large Animal Clinical Sciences, Virginia Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061, USA.
Blikslager
A T
AT
Little
D
D
Howard
R D
RD
Woody
S L
SL
Beex
L M
LM
Crisman
M V
MV
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern / Q000662:veterinary; D003001:Cloning, Molecular; D003085:Colic / Q000209:etiology / Q000662:veterinary; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D015723:Gene Library; D006734:Horse Diseases / Q000235:genetics*; D006736:Horses; D015151:Immunoblotting / Q000662:veterinary; D007413:Intestinal Mucosa / Q000098:blood supply* / Q000378:metabolism; D007511:Ischemia / Q000235:genetics / Q000662:veterinary*; D007583:Jejunum / Q000098:blood supply*; D008297:Male; D012313:RNA / Q000378:metabolism
D012313:RNA