Equine veterinary journal. Supplement2007; (36); 359-364; doi: 10.1111/j.2042-3306.2006.tb05569.x

Gene expression profiling in equine muscle tissues using mouse cDNA microarrays.

Mucher, E
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Jayr, L
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Rossignol, F
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Amiot, F
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Gidrol, X
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Barrey, E

Abstract: Progress could be achieved by using microarrays to understand metabolic adaptations and disorders in equine muscle in response to exercise. Objective: To test the feasibility of using mouse cDNA microarrays to analyse gene expression profile in normal equine muscles. Methods: Muscular biopsies of dorsal gluteus medius and longissimus lumborum were done in 4 healthy Standardbreds. Total RNA was extracted from the muscle samples. The concentration and quality of RNA were measured before and after amplification. Gene expression profiles were measured using mouse cDNA microarrays including 15,264 unique genes representing about 11,000 documented genes. Three hybridisation tests were performed to check interspecificity, reproducibility and to compare gene expression in these muscles. For each test, a dye-swap hybridisation with Cy3 and Cy5 fluoromarkers were done and the gene list filtered according the signal level. Results: According to the specificity test, the mouse cDNA microarrays were correctly hybridised by equine muscle cDNA. All positive control genes (GAPDH, HPRT and beta-Actin) and no negative control gene (yeast, plant) hybridised. The reproducibility test demonstrated a good linearity between the duplicate hybridisations: 99.99% of the significant expressed genes have an expression ratio between 1.4 and 1/1.4 = 0.71. These limits can be considered as the thresholds to qualify as up-regulated (ratio >1.4) or downregulated (ratio <0.71). In the muscle comparison test between gluteus medius vs. longissimus lumborum, 63 genes were found up-regulated and 8 genes down-regulated. The range of gene expression ratios in the gluteus medius was 0.61-8.31 x the longissimus lumborum. This list of modulated genes was classified by functions using a gene ontology data basis. Conclusions: Mouse microarrays could be used to hybridise equine RNA extracted from muscle tissues. For many genes there are large sequence identities that allowed interspecific cDNA hybridisation. The sensitivity of the method allowed quantification of up- and down-regulated genes after applying appropriate filters. Conclusions: Expression profiling could be used to explore the muscle metabolism changes related to exercise, training, pathology and illegal medication in horses.

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Publication Date: 2007-04-04 PubMed ID: 17402448DOI: 10.1111/j.2042-3306.2006.tb05569.xGoogle Scholar: Lookup

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The presented research highlights the application of mouse cDNA microarrays to analyze gene expression in horse muscles, providing valuable information on the metabolic adaptations and complexities of muscle disorders that could potentially occur due to exercise.

Objective and Methodology

The primary objective of this study was to understand the viability of using mouse cDNA microarrays to study gene expression profiles in healthy horse muscles.
The researchers attained muscle biopsies from the dorsal gluteus medius and longissimus lumborum of four healthy Standardbred horses.
Total RNA was extracted from these muscle samples, with the concentration and quality of this RNA measured both pre- and post-amplification.
Gene expression profiles were then examined using mouse cDNA microarrays, incorporating approximately 11,000 recognized genes from 15,264 unique genes.
Three hybridization tests were conducted to determine interspecificity, reproducibility, and to contrast gene expression in these muscles.

Key Results

The mouse cDNA microarrays successfully hybridized with the equine muscle cDNA, making the experimentation process feasible.
All the positive control genes and no negative control genes hybridized, verifying the process’s accuracy and validity.
The reproducibility test showed strong linearity between the duplicate hybridizations, making it possible to effectively quantify up-and down-regulated genes.
When comparing gluteus medius versus longissimus lumborum, the team found 63 up-regulated genes and 8 down-regulated genes. Gene expression ratios ranged notably, suggesting a scope for comprehensive gene expression study.

Conclusions

Mouse cDNA microarrays can effectively hybridize horse RNA from muscle tissues. Since many genes share large sequence similarities, interspecific cDNA hybridization is attainable.
The sensitivity and reliability of this method permit accurate quantification of up- and down-regulated genes following the application of appropriate filters.
Gene expression profiling can be potentially used to investigate muscle metabolism changes in horses linked to exercise, training, pathology, or illegal medication—expanding the scope of equine health and welfare research.

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APA

Mucher E, Jayr L, Rossignol F, Amiot F, Gidrol X, Barrey E. (2007). Gene expression profiling in equine muscle tissues using mouse cDNA microarrays. Equine Vet J Suppl(36), 359-364. https://doi.org/10.1111/j.2042-3306.2006.tb05569.x