Metabonomic study of the biochemical profiles of heterozygous myostatin knockout swine
Jianxiang XU1*, Dengke PAN2*, Wenye Yuan ()1
1 State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100193, China
2 Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
*These authors contributed equally to the work
Correspondence:
Contact: Jianxiang Xu, phone: 13521311702, email:
Abstract Myostatin is a transforming growth factor- family member that normally acts to limit skeletal muscle growth. …
Keywords myostatin, transforming growth factor- family, …
1 Introduction
Metabonomics is... This … transgenesis[1] (superscript). Metabolic changes are directly linked to phenotypic changes, whereas gene expression changes are not; … and physiology.
Nuclear magnetic resonance, liquid chromatography - gas chromatography (LC-MS) ...associated with toxicity[2–8], human disease[9–11], pharmaceutical drugs[12–15], genetic modification[1,16–20].
…
2 Materials and methods
2.1 Animals
All experiments were conducted in accordance with the guidelines of China on the Use and Care of Laboratory Animals. …
2.2 Sample collection
Blood of 26 F1MSTN+/ pigs (Table 1) were obtained …, one part (about 1 mL) was..., Another part (about 500 L) was stored at –80°C until metabonomics analyses.
Table 1 Experimental animas using for this study
Group / Sample / Bodyweight(mean ± SD)/kg / Generation / Description
WT / Male (n = 11) / 114.36 ± 4.02 / F1 / Wild type adult swine (MSTN + /+)
Female (n = 3) / 113.92 ± 5.40
KO / Male (n = 7) / 113.74 ± 5.35 / F1 / Heterozygous MSTN KO adult swine (MSTN+/)
Female (n = 5) / 114.43 ± 5.24
Note: WT, wild type; KO, knock out.
2.3 Sample preparation
Plasma samples (100 L) were extracted using an automated MicroLab STAR® system (Hamilton Company, UT, USA) and …, either LC/MS or GC/MS.
2.4 LC/MS and GC/MS analysis
LC/MS was … methanol both containing 0.1% formic acid, while the basic extracts, which also used water/methanol, contained 6.5 mmol·L–1 ammonium bicarbonate. The MS analysis alternated between MS and data-dependent MS2 scans using dynamic exclusion.
2.5 Data normalization
For studies … normalizing each data point proportionately (Fig. 1). For studies that did not require more than one day of analysis, no normalization was necessary, other than for purposes of data visualization.
Fig. 1 BCAA catabolism. (a) Box plots of the BCAA catabolic key products, there were significant differences (P<0.05) only betweenfemales; (b) BCAA catabolic pathway.
3 Results
3.1 Clinical laboratory analysis
As shown in Table 2,… KO female swine (P < 0.01). ...
…
3.2 Metabolomic analysis
…
3.2.1 Fatty acid metabolism
As shown in Appendix A (Table S1),…
3.2.2 Glucose utilization
…
4 Discussion
4.1 Influencing factors in metabolite profile
Bollard et al. found by metabonomics that time of sampling (day and night) markedly affected the metabolites profiles[26]...
4.2 Metabolomic analysis
…
5 Conclusions
This study demonstrated a few significant differences between WT and MSTN+/swine plasma. …
…
Acknowledgements This work was funded by the key special projects of breeding new varieties of genetically modified organisms in China (2014ZX08012-002). We thank Metabolon, a diagnostic products and services company, for assistance with the metabolic profile and statistical analysis.
Compliance with ethics guidelines Jianxiang Xu, Dengke Pan and Wenye Yuan declare that they have no conflict of interest or financial conflicts to disclose.
All applicable institutional and national guidelines for the care and use of animals were followed.
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