Time course analysis of large-scale gene expression in incised muscle using correspondence analysis.

Tetsuya Horita,Sanae Kanno,Yasuhiro Aoki,Masataka Takamiya,Mamiko Fukuta,Hideaki Kato,Mohammed Hassan Gaballah,Shao Jun Du

doi:10.1371/journal.pone.0230737

Tetsuya Horita, Sanae Kanno + Show 6 more

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https://doi.org/10.1371/journal.pone.0230737

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Abstract

Studying the time course of gene expression in injured skeletal muscle would help to estimate the timing of injuries. In this study, we investigated large-scale gene expression in incision-injured mouse skeletal muscle by DNA microarray using correspondence analysis (CA). Biceps femoris muscle samples were collected 6, 12, and 24 hours after injury, and RNA was extracted and prepared for microarray analysis. On a 2-dimensional plot by CA, the genes (row score coordinate) located farther from each time series (column score coordinate) had more upregulation at particular times. Each gene was situated in 6 subdivided triangular areas according to the magnitude of the relationship of the fold change (FC) value at each time point compared to the control. In each area, genes for which the ratios of two particular FC values were close to 1 were distributed along the two border lines. There was a tendency for genes whose FC values were almost equal to be distributed near the intersection of these 6 areas. Therefore, the gene marker candidates for estimation of the timing of injuries were detectable according to the location on the CA plot. Moreover, gene sets created by a specific gene and its surrounding genes were composed of genes that showed similar or identical fluctuation patterns to the specific gene. In various analyses on these sets, significant gene ontology term and pathway activity may reflect changes in specific genes. In conclusion, analyses of gene sets based on CA plots is effective for investigation of the time-dependent fluctuation in gene expression after injury.

Highlights

The time course of wound healing or the estimation of wound age is one of the most important research subjects in forensic pathology
The animal experiment was approved by the Nagoya City University (NCU) animal ethics committee, and conducted according to the principles of laboratory animal care, and the guidelines for animal experimentation, NCU [10, 11]
The most common pattern was that of insignificant fluctuation throughout the time, which included 22,872 genes or about 40% of all genes

Summary

Introduction

The time course of wound healing or the estimation of wound age is one of the most important research subjects in forensic pathology. A number of research projects have been performed, and the majority of them have dealt with the timing of dermal injuries [1]. Like skin, distributed throughout the whole body, and often affected in cases of fatal and serious injuries such as stab wounds and incisions. Different molecular diagnostic techniques have been used to evaluate the usefulness of many markers for estimating wound age, including cytokines and chemokines.

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