Dynamical Expression of MicroRNA-127-3p in Proliferating and Differentiating C2C12 Cells

MicroRNAs (miRNAs) are highly conserved, short non-coding RNAs that regulate gene expression at the posttranscriptional level. Although many miRNAs are identified in muscles and muscle cells, their individual roles are still not fully understood. In the present study, we investigated a muscle highly-expressed miRNA, miR-127-3p, in C2C12 myoblasts and tissues of goats with different muscle phenotypes (Boer vs Wushan black goats). Our results demonstrated that i) miR-127-3p was extensively expressed in tissues of goats; ii) miR-127-3p was higher expressed in muscle, spleen, heart, and skin in the muscular goats (Boer goats) than the control (Wushan black goats). Then we further characterized the dynamical expression of miR-127-3p, MyoD, MyoG, Myf5, Mef2c, and Myosin in the proliferating and differentiating C2C12 myoblasts at day of 0, 1, 3, 5, and 7 in culture mediums. Especially, we found that miR-127-3p was significantly higher expressed in the proliferating than differentiating cells. Our findings suggest that miR-127-3p probably plays roles in the proliferation and differentiation of myoblasts, which further underlies regulation of muscle phenotype in goats

Histological and transcriptome-wide level characteristics of fetal myofiber hyperplasia during the second half of gestation in Texel and Ujumqin sheep

<p>Abstract</p> <p>Background</p> <p>Whether myofibers increase with a pulsed-wave mode at particular developmental stages or whether they augment evenly across developmental stages in large mammals is unclear. Additionally, the molecular mechanisms of myostatin in myofiber hyperplasia at the fetal stage in sheep remain unknown. Using the first specialized transcriptome-wide sheep oligo DNA microarray and histological methods, we investigated the gene expression profile and histological characteristics of developing fetal ovine longissimus muscle in Texel sheep (high muscle and low fat), as a myostatin model of natural mutation, and Ujumqin sheep (low muscle and high fat). Fetal skeletal muscles were sampled at 70, 85, 100, 120, and 135 d of gestation.</p> <p>Results</p> <p>Myofiber number increased sharply with a pulsed-wave mode at certain developmental stages but was not augmented evenly across developmental stages in fetal sheep. The surges in myofiber hyperplasia occurred at 85 and 120 d in Texel sheep, whereas a unique proliferative surge appeared at 100 d in Ujumqin sheep. Analysis of the microarray demonstrated that immune and hematological systems' development and function, lipid metabolism, and cell communication were the biological functions that were most differentially expressed between Texel and Ujumqin sheep during muscle development. Pathways associated with myogenesis and the proliferation of myoblasts, such as calcium signaling, chemokine (C-X-C motif) receptor 4 signaling, and vascular endothelial growth factor signaling, were affected significantly at specific fetal stages, which underpinned fetal myofiber hyperplasia and postnatal muscle hypertrophy. Moreover, we identified some differentially expressed genes between the two breeds that could be potential myostatin targets for further investigation.</p> <p>Conclusions</p> <p>Proliferation of myofibers proceeded in a pulsed-wave mode at particular fetal stages in the sheep. The myostatin mutation changed the gene expression pattern in skeletal muscle at a transcriptome-wide level, resulting in variation in myofiber phenotype between Texel and Ujumqin sheep during the second half of gestation. Our findings provide a novel and dynamic description of the effect of myostatin on skeletal muscle development, which contributes to understanding the biology of muscle development in large mammals.</p

Optimization of Stope Structure Parameters Based on the Mined Orebody at the Meishan Iron Mine

Based on the engineering background of the Meishan iron mine with sublevel caving (SLC) method, in this work, we adopted the method for identifying the shape of mined orebody (the original location in blasted slice), which analyzed and determined reasonable stope structure parameters. In the field test, the markers were arranged in the blasted slice, the mined orebody was measured by in situ tests, and reliable data were achieved. The shape of the mined orebody was obtained through this test when the width of drift was 6 m. The mined orebody’s shape was compared with the shape of the isolated extraction zone (IEZ), and the difference increased with increasing height. When the stope structural parameters were determined by the mined orebody, the larger the sublevel height was, the smaller the error was, which was compared with the method using ellipsoid arrangement theory to determine the stope structural parameters. Finally, the reasonable stope structure parameters were optimized. The sublevel height was 22 m, and the drift spacing was 20 m

Study on the Process and Mechanism of Slope Failure Induced by Mining under Open Pit Slope: A Case Study from Yanqianshan Iron Mine, China

Mining under an open pit slope results in the collapse and slide of the slope. In this paper, a combination of methods including Google Earth and field investigations is applied to investigate the process of eastern slope failure induced by underground mining in the Yanqianshan Iron Mine over five years. According to the observed ground deformation features, the geomorphic zone of the eastern slope can be divided into four regions (caved rock zone, cracking zone, toppling zone, and sliding zone). Break angles and fracture initiation angles at different times are counted separately. Based on the above work, the process of initiation and development of slope failure is studied. The analysis results show that the process of slope failure could be chronologically divided into three stages. First, a collapse pit, caused by the falling of the overlying strata above the goaf, appeared on the eastern slope. Then, the rock mass around the collapse pit slid into the pit to form a small landslide. Finally, because of mining disturbances and rock creep, a large landslide occurred on the northeastern phyllite slope. The control mechanisms of each failure stage are discussed separately. Finally, the RFPA3D code is employed to simulate the slope failure process under the influence of underground mining. The results are consistent with the field observations, which provided information on deformation failure and the mechanics of the slope that could not be directly observed in the field and deepened the mechanism analysis

A Dynamic Intersecting Arrangement Model Based on Isolated Draw Zones for Stope Structure Optimization during Sublevel Caving Mining

In this paper, stope structure optimization during sublevel caving mining is considered under the condition that the isolated draw zones (IDZs) are nonstandard ellipsoid, which is realized by dynamically adjusting the arrangement of IDZs and quantifying the degree of intersection of IDZs according to an ore profit and loss calculation model. A dynamic intersecting arrangement model based on IDZs was proposed, which can dynamically adjust the sublevel height and drift spacing according to the ore-rock bulk flow parameters, economic indicators, occurrence condition of the ore body, drilling machine, and so forth. Based on the model, the range of drift spacing, the lower volume of crestal residual ore, and the higher volume of mixing waste rock are calculated. By deducing the function of ore profit and loss, a calculation model for ore profit and loss is established to quantify the degree of intersection of IDZs and determine the best stope structure. Using the constructed dynamic intersecting arrangement model, a stope structure of −213 m to −303 m in the Yanqianshan Iron Mine was designed, with a sublevel height of 22.5 m and a drift spacing of 20.5 m. A physical drawing model was designed, and three physical simulation experiment schemes were conducted to compare and analyse the ore loss and dilution of the intersecting arrangement model and the traditional tangent arrangement model. The results showed that the loss rate decreased by 3.66% and the dilution rate increased by only 0.22%, thus verifying the effectiveness and applicability of the model to optimize the stope structure

The microRNA-127-3p directly targeting Vamp2 in C2C12 myoblasts

MicroRNAs (miRNAs) have been reported that can regulate skeletal muscle growth and development. Previously, we demonstrated that miR-127-3p were differently expressed in skeletal muscle and muscle cells. However, the molecular mechanism of miR-127-3p regulation of skeletal myogenesis are not well elucidated. In this study, we transfected miR-127-3p into C2C12 cells, and found miR-127-3p induces myogenesis by targeting Vamp2. Moreover, the regulatory mechanism of Vamp2 in myoblasts proliferation and differentiation was further confirmed. In conclusion, our data providedevidences that miR-127-3p reciprocally regulated myoblasts proliferation and differentiation through directly targeting Vamp2

Identifying Candidate Genes for Litter Size and Three Morphological Traits in Youzhou Dark Goats Based on Genome-Wide SNP Markers

This study aimed to reveal the potential genetic basis for litter size, coat colour, black middorsal stripe and skin colour by combining genome-wide association analysis (GWAS) and selection signature analysis and ROH detection within the Youzhou dark (YZD) goat population (n = 206) using the Illumina GoatSNP54 BeadChip. In the GWAS, we identified one SNP (snp54094-scaffold824-899720) on chromosome 11 for litter size, two SNPs on chromosome 26 (snp11508-scaffold142-1990450, SORCS3) and chromosome 12 (snp55048-scaffold842-324525, LOC102187779) for coat colour and one SNP on chromosome 18 (snp56013-scaffold873-22716, TCF25) for the black middorsal stripe. In contrast, no SNPs were identified for skin colour. In selection signature analysis, 295 significant iHS genomic regions with a mean |iHS| score > 2.66, containing selection signatures encompassing 232 candidate genes were detected. In particular, 43 GO terms and one KEGG pathway were significantly enriched in the selected genes, which may contribute to the excellent environmental adaptability and characteristic trait formation during the domestication of YZD goats. In ROH detection, we identified 4446 ROH segments and 282 consensus ROH regions, among which nine common genes overlapped with those detected using the iHS method. Some known candidate genes for economic traits such as reproduction (TSHR, ANGPT4, CENPF, PIBF1, DACH1, DIS3, CHST1, COL4A1, PRKD1 and DNMT3B) and development and growth (TNPO2, IFT80, UCP2, UCP3, GHRHR, SIM1, CCM2L, CTNNA3 and CTNNA1) were revealed by iHS and ROH detection. Overall, this study is limited by the small population size, which affects the results of GWAS to a certain extent. Nevertheless, our findings could provide the first overview of the genetic mechanism underlying these important traits and provide novel insights into the future conservation and utilisation of Chinese goat germplasm resources

Signals of Ezh2, Src, and Akt Involve in myostatin-Pax7 pathways regulating the myogenic fate determination during the sheep myoblast proliferation and differentiation.

Myostatin and Pax7 have been well documented individually, however, the mechanism by which Myostatin regulates Pax7 is seldom reported. Here, based on muscle transcriptome analysis in Texel (Myostatin mutant) and Ujumqin (wild type) sheep across the five fetal stages, we constructed and examined the Myostatin-Pax7 pathways in muscle. Then we validated the signals by RNAi in the proliferating and differentiating sheep myoblasts in vitro at mRNA, protein, and cell morphological levels. We reveal that Myostatin signals to Pax7 at least through Ezh2, Src, and Akt during the sheep myoblast proliferation and differentiation. Other signals such as p38MAPK, mTOR, Erk1/2, Wnt, Bmp2, Smad, Tgfb1, and p21 are most probably involved in the Myostatin-affected myogenic events. Myostatin knockdown significantly reduces the counts of nucleus and myotube, but not the fusion index of myoblasts during cell differentiation. In addition, findings also indicate that Myostatin is required for normal myogenic differentiation of the sheep myoblasts, which is different from the C2C12 myoblasts. We expand the regulatory network of Myostatin-Pax7 pathways and first illustrate that Myostatin as a global regulator participates in the epigenetic events involved in myogenesis, which contributes to understand the molecular mechanism of Myostatin in regulation of myogenesis