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Comparative Study
. 2011;6(5):e19774.
doi: 10.1371/journal.pone.0019774. Epub 2011 May 26.

Comparative analyses by sequencing of transcriptomes during skeletal muscle development between pig breeds differing in muscle growth rate and fatness

Affiliations
Comparative Study

Comparative analyses by sequencing of transcriptomes during skeletal muscle development between pig breeds differing in muscle growth rate and fatness

Xiao Zhao et al. PLoS One. 2011.

Abstract

Understanding the dynamics of muscle transcriptome during development and between breeds differing in muscle growth is necessary to uncover the complex mechanism underlying muscle development. Herein, we present the first transcriptome-wide longissimus dorsi muscle development research concerning Lantang (LT, obese) and Landrace (LR, lean) pig breeds during 10 time-points from 35 days-post-coitus (dpc) to 180 days-post-natum (dpn) using Solexa/Illumina's Genome Analyzer. The data demonstrated that myogenesis was almost completed before 77 dpc, but the muscle phenotypes were still changed from 77 dpc to 28 dpn. Comparative analysis of the two breeds suggested that myogenesis started earlier but progressed more slowly in LT than in LR, the stages ranging from 49 dpc to 77 dpc are critical for formation of different muscle phenotypes. 595 differentially expressed myogenesis genes were identified, and their roles in myogenesis were discussed. Furthermore, GSK3B, IKBKB, ACVR1, ITGA and STMN1 might contribute to later myogenesis and more muscle fibers in LR than LT. Some myogenesis inhibitors (ID1, ID2, CABIN1, MSTN, SMAD4, CTNNA1, NOTCH2, GPC3 and HMOX1) were higher expressed in LT than in LR, which might contribute to more slow muscle differentiation in LT than in LR. We also identified several genes which might contribute to intramuscular adipose differentiation. Most important, we further proposed a novel model in which MyoD and MEF2A controls the balance between intramuscular adipogenesis and myogenesis by regulating CEBP family; Myf5 and MEF2C are essential during the whole myogenesis process while MEF2D affects muscle growth and maturation. The MRFs and MEF2 families are also critical for the phenotypic differences between the two pig breeds. Overall, this study contributes to elucidating the mechanism underlying muscle development, which could provide valuable information for pig meat quality improvement. The raw data have been submitted to Gene Expression Omnibus (GEO) under series GSE25406.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Morphological variations of longissimus muscle samples during development and between breeds.
LR indicates Landrace and LT indicates Lantang. Numbers (1–10) indicate distinct developmental stages including 35 (1), 49 (2), 63 (3), 77 (4), 91 (5) days post coitus (dpc) and 2 (6), 28 (7), 90 (8), 120 (9), 180 (10) days post natum (dpn). Arrows point to myofibers or muscle fibers, P indicates primary fiber and S indicates secondary fiber. All areas were photographed at a magnification of ×400.
Figure 2
Figure 2. Similarity of transcriptome profiles between 20 skeletal muscle libraries.
(A) Correlation analysis of 20 libraries by Pearson's correlation coefficient. (B) Systematic cluster analysis of 20 libraries by Cluster 3.0 and TreeView software.
Figure 3
Figure 3. The muscle-related biological processes of differentially expressed genes during different muscle development stages.
(A) Muscle development-related GO categories were up-regulated at 49 and 60 dpc and down-regulated at 77 dpc for Landrace. (B) Muscle development-related GO categories were up-regulated at 49 and 60 dpc and down-regulated at 77 dpc for Lantang.
Figure 4
Figure 4. The expression patterns of myogenesis and adipogenesis genes (A, B) and Q-PCR validation of sequencing data (C).
(A) The expression pattern of genes could relate to adipogenesis. The vertical axis indicates the normalized gene expression level in breed or stages; dpc indicates prenatal and dpn indicates postnatal. (B) The expression patterns of MRF and MEF2 families. (C) Validation of sequencing data by Q-PCR. The vertical axis indicates the fold changes of transcript abundance in 20 samples compared to the LR 1. The r value indicates Spearman's Correlation between the two methods.
Figure 5
Figure 5. The muscle developmental genes which were higher expressed in LT than in LR.
(A) These 53 genes were similar expressed with MyoD; (B) These 10 genes were only highly expressed in prenatal LT; (C) GADD45G and FHL1 were peak expressed at 2 dpn and 28 dpn; (D) These 10 genes were similar expressed with MyoD in prenatal, but highly expressed at 2, 28 and 90 dpn of landrace while 90, 120 and 180 dpn of Lantang. (E) The DE genes could relate to muscle development in postnatal between the two breeds.
Figure 6
Figure 6. The muscle developmental genes which were higher expressed in LR than in LT.
(A) These 13 genes were highly expressed at 35 dpc of the two breeds, and then maintained the high expression in prenatal LR but down regulated in prenatal LT; (B) These 7 genes were similar expressed with MEF2D; (C) These 9 genes were highly expressed in prenatal of the two breeds, but higher expressions were showed in LR than in LT; (D) These genes were differential expressed in prenatal between the two breeds, but showed no differences in postnatal; (E) LSM4 and DN123732 were highly expressed in prenatal LR and 2 dpn of LT; (F) These 5 genes were only highly expressed in LR.
Figure 7
Figure 7. The interaction network between the differentially expressed muscle developmental genes.
(A–B) Signal-flow analyses of myogenesis-related genes in LR (A) and LT (B). Yellow dots indicate DE genes and blue dots indicate non-DE genes. Straight lines indicate interaction relationships between genes; area indicates active and flat indicates inhibited. Solid line indicates a direct interaction and dashed line indirect interaction. Value and diameter of line indicates the size of interaction; the larger the values the stronger the regulation.
Figure 8
Figure 8. The molecular regulation of myogenesis and the model for MyoD controls intramuscular adipogenesis and myogenesis.
Red dots indicate the promoting roles of these genes in myogenesis and blank striping indicate the repressing roles. (A) The probable roles of DE genes in regulating myogenesis (B) The novel model for MyoD controls the balance between intramuscular adipogenesis and myogenesis.

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