scholarly journals Analysis of mRNA expression of CNN3, DCN, FBN2, POSTN, SPARC and YWHAQ genes in porcine foetal and adult skeletal muscles

2008 ◽  
Vol 53 (No. 5) ◽  
pp. 181-186 ◽  
Author(s):  
K. Bílek ◽  
A. Knoll ◽  
A. Stratil ◽  
K. Svobodová ◽  
P. Horák ◽  
...  
Keyword(s):  
Skeletal Muscles ◽  
Muscle Growth ◽  
Biceps Femoris ◽  
Subtractive Hybridization ◽  
Postnatal Growth ◽  
Prenatal Development ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Muscle Biology ◽  
Fetal Muscle

Skeletal muscle growth is determined by the number of prenatally formed fibres and by the degree of their postnatal hypertrophy; i.e. prenatal development may influence the postnatal growth. Suppression subtractive hybridization (SSH) was used to identify genes more expressed in fetal hind limb muscles of Piétrain pigs (44 days of gestation) compared to the adult biceps femoris. Six potential functional candidate genes (<i>CNN3</i>, <i>DCN</i>, <i>FBN2</i>, <i>POSTN</i>, <i>SPARC</i> and <i>YWHAQ</i>) were selected to verify the SSH results using real-time RT-PCR. Expression levels of the studied genes were significantly higher (<i>P</i>< 0.05) in the fetal muscle compared to the adult muscle. <i>FBN</i>2 and <i>POSTN</i> exhibited the highest mRNA levels (mean relative ratios were 182.7 and 121.6, respectively). The studied genes may play an important role in muscle biology and may be candidates for muscling traits.

scholarly journals Expression profiles of myostatin and calpastatin genes and analysis of shear force and intramuscular fat content of yak longissimus muscle

2011 ◽  
Vol 56 (No. 12) ◽  
pp. 545-550 ◽  
Author(s):  
Y.C. Zheng ◽  
Y.Q. Lin ◽  
Y. Yue ◽  
Y.O. Xu ◽  
S.Y. Jin
Keyword(s):  
Expression Profiles ◽  
Muscle Growth ◽  
Intramuscular Fat ◽  
Mrna Levels ◽  
Longissimus Muscle ◽  
Rt Pcr ◽  
Yellow Cattle ◽  
Significant Difference ◽  
Different Ages ◽  
Imf Content

The main objective of this study was to reveal the expression profiles of two negative regulators, myostatin (MSTN) and calpastatin (CAST)genes, of skeletal muscle growth in highland yaks (Bos grunniens). mRNA levels of both genes were quantified in different yak tissues by semi-quantitative RT-PCR to reveal the tissue expression pattern, and real-time quantitative RT-PCR was employed to compare the mRNA levels of MSTN and CAST in longissimus muscles of yaks at different ages and adult Yellow cattle. Intramuscular fat (IMF) content, tenderness and pH of longissimus muscle of yaks at different ages and of adult Yellow cattle were also measured. The results showed that MSTN and CAST expressions have tissue specificity and both exhibited a high level in longissimus muscle and a low level in adipose tissue. Yak calves had lower mRNA levels of both MSTN and CAST in longissimus muscle compared with adult yaks. The analysis of meat quality traits of longissimus muscle showed that the shear forces of raw longissimus muscle of yak calves were significantly lower than those of adult yaks and Yellow cattle, no significant difference was found between adult yaks and Yellow cattle of similar age. IMF content in longissimus muscle was lower in yaks than in Yellow cattle. Although yaks were smaller in body size than Yellow cattle, adult yaks showed lower levels of MSTN and similar level of CAST mRNA in longissimus muscle compared to Yellow cattle. These data indicate that the expression of both MSTN and CAST in longissimus muscle differs between adult yaks and yak calves, and the yak longissimus muscle shows a lower IMF content compared to cattle. &nbsp;

scholarly journals Fetal ovine skeletal and cardiac muscle transcriptomics are differentially altered by increased maternal cortisol during gestation

2020 ◽  
Vol 52 (4) ◽  
pp. 178-190 ◽  
Author(s):  
Serene Joseph ◽  
Bryan Alava ◽  
Andrew Antolic ◽  
Elaine M. Richards ◽  
Charles E. Wood ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cytochrome C ◽  
Interventricular Septum ◽  
Biceps Femoris ◽  
Mitochondrial Metabolism ◽  
Mitochondrial Activity ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Mt Dna ◽  
Biceps Femoris Muscle

We have previously found that in utero exposure to excess maternal cortisol (1 mg/kg/day) in late gestation increases the incidence of stillbirth during labor and produces fetal bradycardia at birth. In the interventricular septum, mitochondrial DNA (mt-DNA) was decreased, and transcriptomics and metabolomics were consistent with altered mitochondrial metabolism. The present study uses transcriptomics to model effects of increased maternal cortisol on fetal biceps femoris. Transcriptomic modeling revealed that pathways related to mitochondrial metabolism were downregulated, whereas pathways for regulation of reactive oxygen species and activation of the apoptotic cascade were upregulated. Mt-DNA and the protein levels of cytochrome C were significantly decreased in the biceps femoris. RT-PCR validation of the pathways confirmed a significant decrease in SLC2A4 mRNA levels and a significant increase in PDK4, TXNIP, ANGPTL4 mRNA levels, suggesting that insulin sensitivity of the biceps femoris muscle may be reduced in cortisol offspring. We also tested for changes in gene expression in diaphragm by rt-PCR. PDK4, TXNIP, and ANGPTL4 mRNA were also increased in the diaphragm, but SLC2A4, cytochrome C protein, and mt-DNA were unchanged. Comparison of the change in gene expression in biceps femoris to that in cardiac interventricular septum and left ventricle showed few common genes and little overlap in specific metabolic or signaling pathways, despite reduction in mt-DNA in both heart and biceps femoris. Our results suggest that glucocorticoid exposure alters expression of nuclear genes important to mitochondrial activity and oxidative stress in both cardiac and skeletal muscle tissues, but that these effects are tissue-specific.

Isolation of genes involved in pancreas regeneration by subtractive hybridization

2010 ◽  
Vol 391 (9) ◽  
Author(s):  
Jong-Ho Choi ◽  
Min-Young Lee ◽  
Yoolee Kim ◽  
Jeong-Yun Shim ◽  
Sang-Moon Han ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Embryonic Stem ◽  
Subtractive Hybridization ◽  
Marker Genes ◽  
Mrna Levels ◽  
Genetic Therapy ◽  
Rt Pcr ◽  
Β Cells ◽  
Paracrine Factors ◽  
Pancreas Regeneration

AbstractThe deterioration of β cells in the pancreas is a crucial factor in the progression of diabetes mellitus; therefore, the recovery of β cells is of vital importance for effective diabetic therapeutic strategies. Partially pancreatectomized rats have been used for the investigation of pancreatic regeneration. Because it was determined that tissue extract from the partially-dissected pancreas induces pancreatic differentiation in embryonic stem cells, paracrine factors were thought to be involved in the regeneration. In this study, we screened for genes that had higher mRNA levels 2 days after 60%-pancreatectomy. The genes were isolated using subtractive hybridization and DNA sequencing. Twelve genes (adipsin,Aplp2,Clu,Col1a2,Glul,Krt8,Lgmn, LOC299907, LOC502894,Pla2g1b,Reg3αandXbp1) were identified, and RT-PCR and real-time PCR analyses were performed to validate their expression levels. Among the genes identified, three genes (Glul,LgmnandReg3a) were selected for further analyses. Assays revealed thatGlulandReg3αenhance cell growth.Glul,LgmnandReg3αchange the expression level of islet marker genes, whereNEUROD,NKX2.2,PAX4andPAX6are up-regulated andsomatostatinis down-regulated. Thus, we believe thatGlul,LgmnandReg3acan serve as novel targets in diabetes mellitus genetic therapy.

Negative Correlation Between Serum Levels of Homocysteine and Apolipoprotein M

2019 ◽  
Vol 19 (2) ◽  
pp. 120-126
Author(s):  
J. Wei ◽  
Y. Yu ◽  
Y. Feng ◽  
J. Zhang ◽  
Q. Jiang ◽  
...  
Keyword(s):  
Negative Correlation ◽  
Hepg2 Cells ◽  
Serum Levels ◽  
Significant Negative Correlation ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Apolipoprotein M ◽  
Protein Mass ◽  
Protein Levels ◽  
Phosphoinositide 3 Kinase

Background: Homocysteine (Hcy) has been suggested as an independent risk factor for atherosclerosis. Apolipoprotein M (apoM) is a constituent of the HDL particles. The goal of this study was to examine the serum levels of homocysteine and apoM and to determine whether homocysteine influences apoM synthesis. Methods: Serum levels of apoM and Hcy in 17 hyperhomocysteinemia (HHcy) patients and 19 controls were measured and their correlations were analyzed. Different concentrations of homocysteine (Hcy) and LY294002, a specific phosphoinositide 3- kinase (PI3K) inhibitor, were used to treat HepG2 cells. The mRNA levels were determined by RT-PCR and the apoM protein mass was measured by western blot. Results: We found that decreased serum apoM levels corresponded with serum HDL levels in HHcy patients, while the serum apoM levels showed a statistically significant negative correlation with the serum Hcy levels. Moreover, apoM mRNA and protein levels were significantly decreased after the administration of Hcy in HepG2 cells, and this effect could be abolished by addition of LY294002. Conclusions: resent study demonstrates that Hcy downregulates the expression of apoM by mechanisms involving the PI3K signal pathway.

scholarly journals Phenotypical and Functional Characteristics of Human Regulatory T Cells during Ex Vivo Maturation from CD4+ T Lymphocytes

2021 ◽  
Vol 11 (13) ◽  
pp. 5776
Author(s):  
Varvara G. Blinova ◽  
Natalia S. Novachly ◽  
Sofya N. Gippius ◽  
Abdullah Hilal ◽  
Yulia A. Gladilina ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Ex Vivo ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Inflammatory Reactions ◽  
Effector Cells ◽  
Cycle Regulation ◽  
Further Development

Regulatory T cells (Tregs) participate in the negative regulation of inflammatory reactions by suppressing effector cells. In a number of autoimmune disorders, the suppressive function and/or the number of Tregs is compromised. The lack of active functioning Tregs can be restored with adoptive transfer of expanded ex vivo autologous Tregs. In our study, we traced the differentiation and maturation of Tregs CD4+CD25+FoxP3+CD127low over 7 days of cultivation from initial CD4+ T cells under ex vivo conditions. The resulting ex vivo expanded cell population (eTregs) demonstrated the immune profile of Tregs with an increased capacity to suppress the proliferation of target effector cells. The expression of the FoxP3 gene was upregulated within the time of expansion and was associated with gradual demethylation in the promotor region of the T cell-specific demethylation region. Real-time RT-PCR analysis revealed changes in the expression profile of genes involved in cell cycle regulation. In addition to FOXP3, the cells displayed elevated mRNA levels of Ikaros zinc finger transcription factors and the main telomerase catalytic subunit hTERT. Alternative splicing of FoxP3, hTERT and IKZF family members was demonstrated to be involved in eTreg maturation. Our data indicate that expanded ex vivo eTregs develop a Treg-specific phenotype and functional suppressive activity. We suggest that eTregs are not just expanded but transformed cells with enhanced capacities of immune suppression. Our findings may influence further development of cell immunosuppressive therapy based on regulatory T cells.

scholarly journals Quantitative assessment of CYP11B1 and CYP11B2 expression in aldosterone-producing adenomas

2002 ◽  
pp. 795-802 ◽  
Author(s):  
F Fallo ◽  
V Pezzi ◽  
L Barzon ◽  
P Mulatero ◽  
F Veglio ◽  
...  
Keyword(s):  
Real Time ◽  
Secretory Activity ◽  
Zona Glomerulosa ◽  
Zona Fasciculata ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Pathophysiological Role ◽  
Aldosterone Production

BACKGROUND: The presence and pathophysiological role of CYP11B1 (11beta-hydroxylase) gene in the zona glomerulosa of human adrenal cortex is still controversial. METHODS: In order to specifically quantify CYP11B1, CYP11B2 (aldosterone synthase) and CYP17(17alpha-hydroxylase) mRNA levels, we developed a real-time RT-PCR assay and examined the expression in a series of adrenal tIssues, including six normal adrenals from patients adrenalectomized for renal cancer and twelve aldosterone-producing adenomas (APA) from patients with primary aldosteronism. RESULTS: CYP11B1 mRNA levels were clearly detected in normal adrenals, which comprised both zona glomerulosa and fasciculata/reticularis cells, but were also measured at a lower range (P<0.05) in APA. The levels of CYP11B2 mRNA were lower (P<0.005) in normal adrenals than in APA. CYP17 mRNAlevels were similar in normal adrenals and in APA. In patients with APA, CYP11B2 and CYP11B1 mRNA levels were not correlated either with basal aldosterone or with the change from basal aldosterone in response to posture or to dexamethasone. No correlation between CYP11B1 mRNA or CYP11B2 mRNA and the percentage of zona fasciculata-like cells was observed in APA. CONCLUSIONS: Real-time RT-PCR can be reliably used to quantify CYP11B1 and CYP11B2 mRNA levels in adrenal tIssues. Expression of CYP11B1 in hyperfunctioning zona glomerulosa suggests an additional formation of corticosterone via 11beta-hydroxylase, providing further substrate for aldosterone biosynthesis. CYP11B1 and CYP11B2 mRNA levels in APA are not related to the in vivo secretory activity of glomerulosa cells, where post-transcriptional factors might ultimately regulate aldosterone production.

Relative Levels of mRNA Encoding Enamel Proteins in Enamel Organ Epithelia and Odontoblasts

2003 ◽  
Vol 82 (12) ◽  
pp. 982-986 ◽  
Author(s):  
T. Nagano ◽  
S. Oida ◽  
H. Ando ◽  
K. Gomi ◽  
T. Arai ◽  
...  
Keyword(s):  
Tooth Enamel ◽  
Structural Proteins ◽  
Enamel Organ ◽  
Maturation Stage ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Enamel Protein ◽  
Enamel Proteins ◽  
Enamel Layer

Amelogenin, enamelin, sheathlin (ameloblastin/ amelin), enamelysin (MMP-20), and KLK4 (EMSP-1) are the major structural proteins and proteinases in developing tooth enamel. Recently, odontoblasts were reported to express amelogenin, the most abundant enamel protein. In this study, we hypothesized that odontoblasts express all enamel proteins and proteases, and we measured their relative mRNA levels in enamel organ epithelia and odontoblasts associated with porcine secretory- and maturation-stage enamel by RT-PCR, using a LightCycler instrument. The results showed that amelogenin mRNA in secretory-stage EOE is 320-fold higher than in odontoblasts beneath secretory-stage enamel, and over 20,000-fold higher than in odontoblasts under maturation-stage enamel. Similar results were obtained for enamelin and sheathlin. Enamelysin mRNA levels were equivalent in these two tissues, while KLK4 mRNA was higher in odontoblasts than in secretory-stage EOE. These results support the conclusion that odontoblasts are involved in the formation of the enamel layer adjacent to enamel-dentin junction.

scholarly journals Steroidogenic enzymes mRNA expression profile and steroids production in bovine theca cells cultured in vitro and stimulated with angiotensin II

2015 ◽  
Vol 45 (4) ◽  
pp. 704-710 ◽  
Author(s):  
Melânia Lazzari Rigo ◽  
Andressa Minussi Pereira Dau ◽  
Werner Giehl Glanzner ◽  
Manoel Martins ◽  
Renato Zanella ◽  
...  
Keyword(s):  
High Performance ◽  
Culture Media ◽  
Mrna Levels ◽  
Ang Ii ◽  
Rt Pcr ◽  
Steroidogenic Enzymes ◽  
Theca Cells ◽  
Mrna Profile ◽  
Dose Response Effect

The main objective of this study was to detect the steroidogenic effects of Ang II in bovine theca cells in vitro. Bovine theca cells were obtained from follicles (larger than 10mm of diameter) collected from a local abattoir and submitted to different treatments in a sequence of experiments. In experiment 1, CYP17A1 mRNA profile was evaluated in LH- (10ng ml-1) and Ang II-treated (0.1µM) theca cells. In experiment 2, a dose-response effect of Ang II (0.001; 0.1 e 10µM) plus insulin (100ng ml-1) and LH (100ng ml-1) was evaluated on steroidogenesis of bovine theca cells. Experiment 3 explored the effects of saralasin (an antagonist of Ang II receptors) on steroid production and steroidogenic enzymes regulation in theca cells. After 24 hours, culture media from experiments 2 and 3 was collected to evaluate testosterone and androstenedione levels by High-Performance Liquid Chromatography. In parallel, mRNA levels of key steroidogenic enzymes (HSD3B2, CYP11A1, CYP17A1) and STAR were assessed by RT-PCR. There was no difference in testosterone and androstenedione production between treated and controls groups, as well as in mRNA levels of the evaluated genes. In conclusion, the results suggest that Ang II does not regulate steroidogenesis in bovine theca cells

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