Gene and protein expression changes in response to normoxic perfusion in mouse hearts.
BACKGROUND
Although crystalloid-perfused isolated heart models are widely used in cardiovascular research, there are several limitations of these techniques. Changes in cardiac gene expression pattern due to normoxic perfusion itself have not been studied, despite its potential importance to provide useful information on limitations of this model. Therefore, here we investigated the time-dependent effect of normoxic, normothermic perfusion on global gene expression at mRNA and protein levels.


METHODS
Hearts from male CFLP mice were perfused according to the Langendorff technique. We assessed relative gene expression changes by DNA microarray analysis of 8000 genes after 0, 60 and 120 min perfusion.


RESULTS
Twelve genes exhibited significant up-regulation and 27 showed repression in hearts perfused for 60 or 120 min as compared to 0 min controls. Expression changes of 17 selected genes were verified and an additional 19 genes were examined by real-time quantitative PCR. Genes with altered expression included those coding for Creatin kinase, Lactate dehydrogenase, Voltage-dependent anion channel 1, a Disintegrin and Metalloprotease domain 3, Integrin alpha 7, Long-chain acyl-CoA dehydrogenase, Casein kinase II, Ketohexokinase, Chloride ion current inducer protein, Matrix metalloproteinase 2 and 9, Superoxide dismutases and Nitric oxide synthases, etc.


CONCLUSIONS
Our results show that normoxic crystalloid perfusion itself results in time-dependent changes in cardiac gene expression which should be considered when designing ex vivo perfusion protocols in the mouse heart to mimic cardiac pathologies as many of these genes have been suspected to influence several cardiovascular diseases.
Faragó
Nóra
N
Laboratory of Functional Genomics, Biological Research Center, Hungarian Academy of Sciences, H-6726, Szeged, Hungary.
Kocsis
Gabriella F
GF
Fehér
Liliána Z
LZ
Csont
Tamás
T
Hackler
László
L
Varga-Orvos
Zoltán
Z
Csonka
Csaba
C
Kelemen
János Z
JZ
Ferdinandy
Péter
P
Puskás
László G
LG
D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009206:Myocardium / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010100:Oxygen / Q000378:metabolism*; D010477:Perfusion; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014158:Transcription, Genetic
D012333:RNA, Messenger; D007770:L-Lactate Dehydrogenase; D003402:Creatine Kinase; D010100:Oxygen