scholarly journals The Impact of Polyamine Precursors, Polyamines, and Steroid Hormones on Temporal Messenger RNA Abundance in Bovine Satellite Cells Induced to Differentiate

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 764
Author(s):  
Caleb C. Reichhardt ◽  
Lillian L. Okamoto ◽  
Laura A. Motsinger ◽  
Brian P. Griffin ◽  
Gordon K. Murdoch ◽  
...  
Keyword(s):  
Messenger Rna ◽  
Myogenic Differentiation ◽  
Horse Serum ◽  
Biosynthesis Pathway ◽  
Polyamine Biosynthesis ◽  
Trenbolone Acetate ◽  
Steroidal Hormones ◽  
Myogenic Factor ◽  
The Impact ◽  
Anabolic Implants

Emerging research suggests that hormones found in anabolic implants interact with polyamine biosynthesis. The objective of this study was to determine the effects of steroidal hormones, polyamines and polyamine precursors on bovine satellite cell (BSC) differentiation and polyamine biosynthesis temporally. Primary BSCs were induced to differentiate in 3% horse serum (CON) and treated with 10 nM trenbolone acetate (TBA), 10 nM estradiol (E2), 10 nM TBA and 10 nM E2, 10 mM methionine, 8 mM ornithine, 2 mM putrescine, 1.5 mM spermidine, or 0.5 mM spermine. Total mRNA was isolated 0, 2, 4, 8, 12, 24, and 48 h post-treatment. Abundance of mRNA for genes associated with induction of BSC differentiation: paired box transcription factor 7, myogenic factor 5, and myogenic differentiation factor 1 and genes in the polyamine biosynthesis pathway: ornithine decarboxylase and S-adenosylmethionine—were analyzed. Overall, steroidal hormones did not impact (p > 0.05) mRNA abundance of genes involved in BSC differentiation, but did alter (p = 0.04) abundance of genes involved in polyamine biosynthesis. Polyamine precursors influenced (p < 0.05) mRNA of genes involved in BSC differentiation. These results indicate that polyamine precursors and polyamines impact BSC differentiation and abundance of mRNA involved in polyamine biosynthesis, while steroidal hormones altered the mRNA involved in polyamine biosynthesis.

scholarly journals PSIX-28 Examining the effects of estradiol, trenbolone acetate, or polyamines on bovine satellite cell differentiation

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 333-333
Author(s):  
Lillian L Okamoto ◽  
Caleb C Reichhardt ◽  
Brian Griffin ◽  
Laura A Smith ◽  
Gordon Murdoch ◽  
...  
Keyword(s):  
Satellite Cell ◽  
Myogenic Differentiation ◽  
Horse Serum ◽  
Muscle Growth ◽  
Post Treatment ◽  
Growth Promoters ◽  
Trenbolone Acetate ◽  
Physiological Pathways ◽  
Potent Growth ◽  
Anabolic Implants

Abstract The mechanism through which anabolic implants improve skeletal muscle growth of beef animals is incompletely understood. Polyamines (PA) are bioactive amino acid derivatives that act as potent growth stimulants. The objective of this study was to determine effects of anabolic implants, PA and their precursors on bovine satellite cell (BSC) differentiation. Primary BSC were cultured to approximately 80% confluency, at which time they were induced to differentiate in the presence of 3% horse serum (Con) and treated with 10nM TBA, 10 nM E2, or 10nM TBA and 10 nM E2 (ETBA), 10 mM methionine (Met), 8 mM ornithine (Orn), 2 mM putrescine (Put), 1.5 mM spermidine (Spd) or 0.5 mM spermine (Spe). Total mRNA was isolated 0, 2, 4, 8, 12, 24, or 48 h post-treatment and abundance of paired box transcription factor 7 (Pax7) and myogenic differentiation factor (MyoD) were analyzed. Treatment with the hormones (TBA, E2, or ETBA) and PA (Orn, Put, Spd, and Spe) increased (P &lt; 0.05) abundance of MyoD 4 h post-treatment when compared to Con cultures. However, 24 h post-treatment, MyoD abundance was decreased in the presence of hormone treatments when compared to the Con, while the PA treatments increased (P &lt; 0.05) abundance of MyoD when compared to the Con cultures. Treatment with either the hormones or PA had no effect (P &gt; 0.05) on Pax7 abundance at 2, 4, 8, 12, 24, or 48 h post-treatment when compared to Con cultures (P &gt; 0.05). These results indicate that treatment with PA or hormones increases abundance of MyoD, though temporally different indicating that these two classes of growth promoters impact differentiation via alternate physiological pathways. Additional research is underway in order to determine the effects of both PA and hormones on differentiation of primary BSC.

scholarly journals Loss of IGF-IEa or IGF-IEb Impairs Myogenic Differentiation

Endocrinology ◽  
2011 ◽  
Vol 152 (5) ◽  
pp. 1923-1934 ◽  
Author(s):  
Ronald W. Matheny ◽  
Bradley C. Nindl
Keyword(s):  
Splice Variants ◽  
Myogenic Differentiation ◽  
Horse Serum ◽  
Myoblast Differentiation ◽  
Proliferation And Differentiation ◽  
Igf I ◽  
Simultaneous Loss ◽  
Functional Relevance ◽  
Free Media ◽  
The Impact

Actions of protein products resulting from alternative splicing of the Igf1 gene have received increasing attention in recent years. However, the significance and functional relevance of these observations remain poorly defined. To address functions of IGF-I splice variants, we examined the impact of loss of IGF-IEa and IGF-IEb on the proliferation and differentiation of cultured mouse myoblasts. RNA interference-mediated reductions in total IGF-I, IGF-IEa alone, or IGF-IEb alone had no effect on cell viability in growth medium. However, cells deficient in total IGF-I or IGF-IEa alone proliferated significantly slower than control cells or cells deficient in IGF-IEb in serum-free media. Simultaneous loss of both or specific loss of either splice variant significantly inhibited myosin heavy chain (MyHC) immunoreactivity by 70–80% (P &lt; 0.01) under differentiation conditions (48 h in 2% horse serum) as determined by Western immunoblotting. This loss in protein was associated with reduced MyHC isoform mRNAs, because reductions in total IGF-I or IGF-IEa mRNA significantly reduced MyHC mRNAs by approximately 50–75% (P &lt; 0.05). Loss of IGF-IEb also reduced MyHC isoform mRNA significantly, with the exception of Myh7, but to a lesser degree (∼20–40%, P &lt; 0.05). Provision of mature IGF-I, but not synthetic E peptides, restored Myh3 expression to control levels in cells deficient in IGF-IEa or IGF-IEb. Collectively, these data suggest that IGF-I splice variants may regulate myoblast differentiation through the actions of mature IGF-I and not the E peptides.

scholarly journals Resources, Production Scales and Time Required for Producing RNA Vaccines for the Global Pandemic Demand

Vaccines ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 3
Author(s):  
Zoltán Kis ◽  
Cleo Kontoravdi ◽  
Robin Shattock ◽  
Nilay Shah
Keyword(s):  
Production Process ◽  
Messenger Rna ◽  
Vaccination Campaign ◽  
Vaccine Dose ◽  
Economic Feasibility ◽  
Small Scale ◽  
Production Scale ◽  
Working Volume ◽  
Global Demand ◽  
The Impact

To overcome pandemics, such as COVID-19, vaccines are urgently needed at very high volumes. Here we assess the techno-economic feasibility of producing RNA vaccines for the demand associated with a global vaccination campaign. Production process performance is assessed for three messenger RNA (mRNA) and one self-amplifying RNA (saRNA) vaccines, all currently under clinical development, as well as for a hypothetical next-generation saRNA vaccine. The impact of key process design and operation uncertainties on the performance of the production process was assessed. The RNA vaccine drug substance (DS) production rates, volumes and costs are mostly impacted by the RNA amount per vaccine dose and to a lesser extent by the scale and titre in the production process. The resources, production scale and speed required to meet global demand vary substantially in function of the RNA amount per dose. For lower dose saRNA vaccines, global demand can be met using a production process at a scale of below 10 L bioreactor working volume. Consequently, these small-scale processes require a low amount of resources to set up and operate. RNA DS production can be faster than fill-to-finish into multidose vials; hence the latter may constitute a bottleneck.

scholarly journals Neurodegenerative diseases: a hotbed for splicing defects and the potential therapies

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Dunhui Li ◽  
Craig Stewart McIntosh ◽  
Frank Louis Mastaglia ◽  
Steve Donald Wilton ◽  
May Thandar Aung-Htut
Keyword(s):  
Alternative Splicing ◽  
Neurodegenerative Diseases ◽  
Messenger Rna ◽  
Muscular Atrophy ◽  
Single Gene ◽  
Synaptic Function ◽  
Coding Regions ◽  
Splicing Defects ◽  
The Impact ◽  
Splicing Patterns

AbstractPrecursor messenger RNA (pre-mRNA) splicing is a fundamental step in eukaryotic gene expression that systematically removes non-coding regions (introns) and ligates coding regions (exons) into a continuous message (mature mRNA). This process is highly regulated and can be highly flexible through a process known as alternative splicing, which allows for several transcripts to arise from a single gene, thereby greatly increasing genetic plasticity and the diversity of proteome. Alternative splicing is particularly prevalent in neuronal cells, where the splicing patterns are continuously changing to maintain cellular homeostasis and promote neurogenesis, migration and synaptic function. The continuous changes in splicing patterns and a high demand on many cis- and trans-splicing factors contribute to the susceptibility of neuronal tissues to splicing defects. The resultant neurodegenerative diseases are a large group of disorders defined by a gradual loss of neurons and a progressive impairment in neuronal function. Several of the most common neurodegenerative diseases involve some form of splicing defect(s), such as Alzheimer’s disease, Parkinson’s disease and spinal muscular atrophy. Our growing understanding of RNA splicing has led to the explosion of research in the field of splice-switching antisense oligonucleotide therapeutics. Here we review our current understanding of the effects alternative splicing has on neuronal differentiation, neuronal migration, synaptic maturation and regulation, as well as the impact on neurodegenerative diseases. We will also review the current landscape of splice-switching antisense oligonucleotides as a therapeutic strategy for a number of common neurodegenerative disorders.

Effect of Substrate Elasticity on In Vitro Aging of Human Mesenchymal Stem Cells

2012 ◽  
Vol 1498 ◽  
pp. 39-45
Author(s):  
Courtney E. LeBlon ◽  
Caitlin R. Fodor ◽  
Tony Zhang ◽  
Xiaohui Zhang ◽  
Sabrina S. Jedlicka
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Elastic Modulus ◽  
Myogenic Differentiation ◽  
Human Mesenchymal Stem Cells ◽  
Immunofluorescent Staining ◽  
Differentiation Potential ◽  
Gene And Protein Expression ◽  
The Impact

ABSTRACTHuman mesenchymal stem cells (hMSCs) were routinely cultured on tissue-culture polystyrene (TCPS) to investigate the in vitro aging and cell stiffening. hMSCs were also cultured on thermoplastic polyurethane (TPU), which is a biocompatible polymer with an elastic modulus of approximately 12.9MPa, to investigate the impact of substrate elastic modulus on cell stiffening and differentiation potential. Cells were passaged over several generations on each material. At each passage, cells were subjected to osteogenic and myogenic differentiation. Local cell elastic modulus was measured at every passage using atomic force microscopy (AFM) indentation. Gene and protein expression was examined using qRT-PCR and immunofluorescent staining, respectively, for osteogenic and myogenic markers. Results show that the success of myogenic differentiation is highly reliant on the elastic modulus of the undifferentiated cells. The success of osteogenic differentiations is most likely somewhat dependent on the cell elastic modulus, as differentiations were more successful in earlier passages, when cells were softer.

scholarly journals The Impact of Dietary Compounds in Functional Foods on MicroRNAs Expression

2021 ◽  
Author(s):  
Wittaya Chaiwangyen
Keyword(s):  
Messenger Rna ◽  
Positive Impact ◽  
Biologically Active ◽  
Transcriptional Gene Silencing ◽  
Cellular Mirnas ◽  
Rna Molecules ◽  
Post Transcriptional Gene Silencing ◽  
Molecular Machinery ◽  
Different Origins ◽  
The Impact

MicroRNAs (miRNAs) are a class of non-coding endogenous RNA molecules that are involved in post-transcriptional gene silencing via binding to their target messenger RNA, leading to mRNA degradation or translational repression. MicroRNAs can be modulated by several factors including hormones, transcription factors, and dietary compounds. These biologically active compounds have positive impact on the progression of human pathology including non-communicable diseases, which indicating that administration of diet may have potential as therapeutic agents in modulating the risk of chronic diseases. Interestingly, evidence emerging in recent years suggests that dietary miRNAs can be absorbed in human circulation, modulated human gene expression and biological functions. The exploitation of the miRNA functioning within different origins, cellular miRNAs and dietary miRNAs will help us to understand the molecular machinery as well as the regulatory mechanisms involved in fundamentally important biological processes. Therefore, this knowledge may be applied of natural bioactive compounds in preventive or therapeutic approaches.

scholarly journals Myogenic differentiation potential of chicken mesenchymal stem cells from bone marrow

2021 ◽  
Author(s):  
Zhen Zhou ◽  
Changbin Zhao ◽  
Bolin Cai ◽  
Manting Ma ◽  
Shaofen Kong ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Muscle Development ◽  
Myogenic Differentiation ◽  
Horse Serum ◽  
Growth Characteristic ◽  
Normal Growth ◽  
Receptor Interaction ◽  
Differentiation Potential ◽  
Differential Adhesion

Abstract Background: Mesenchymal stem cells (MSCs) have the potential to multilineage differentiation, which can be used for a good model to provide critical insight of chicken muscle development. Differential adhesion method is one of the commonest methods to isolate MSCs based on the ability of plastic adhesion. 5-azacytidine (5-Aza), dexamethasone (DXMS), hydrocortisone (HC) and horse serum had been proved the potential to induce the myogenic differentiation of MSCs. However, the myogenic differentiation of MSCs is still poorly understood in chicken. In present study, we isolated chicken mesenchymal stem cells (cMSCs) from bone using 4-hour differential adhesion method and analyzed the myogenic effect of cMSCs treated with different method based on 5-Aza, DXMS, HC and horse serum.Results: cMSCs isolated by 4-hour differential adhesion method expressed MSCs special surface markers and presented normal growth characteristic. cMSCs showed great potential of myogenic differentiation by the treatment of 5-Aza and horse serum. RNA-sequence, GO and KGEE enrichment analysis revealed that this effect might be based on demethylation of 5-Aza and ECM-receptor interaction, focal adhesion, PI3K-Akt, p53, TGF-beta signaling pathways. Moreover, DXMS, HC and horse serum also presented potential of myogenic differentiation, but the effect was not as good as 5-Aza and horse serum method.Conclusions: cMSCs showed potential of myogenic differentiation by the treatment of 5-Aza and horse serum or DXMS, HC and horse serum.

scholarly journals Scots pine aminopropyltransferases shed new light on evolution of the polyamine biosynthesis pathway in seed plants

2018 ◽  
Vol 121 (6) ◽  
pp. 1243-1256 ◽  
Author(s):  
Jaana Vuosku ◽  
Katja Karppinen ◽  
Riina Muilu-Mäkelä ◽  
Tomonobu Kusano ◽  
G H M Sagor ◽  
...  
Keyword(s):  
Scots Pine ◽  
Seed Plants ◽  
Biosynthesis Pathway ◽  
Polyamine Biosynthesis

scholarly journals PSXIV-32 Oleic acid stimulate the formation of adipocyte-like cells from bovine satellite cells via G Protein Coupled Receptor 43 and CCAAT/Enhancer Binding Protein Beta

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 308-309
Author(s):  
Jongkyoo Kim ◽  
Ki Yong Chung ◽  
Bradley J Johnson ◽  
Stephen B Smith
Keyword(s):  
Gene Expression ◽  
Oleic Acid ◽  
Satellite Cells ◽  
Myogenic Differentiation ◽  
Adipogenic Differentiation ◽  
Horse Serum ◽  
Enhancer Binding Protein ◽  
Mrna Gene ◽  
Fetal Bovine ◽  
Mrna Gene Expression

Abstract Numerous physiological and pathological processes are controlled by free fatty acids (FFAs), which act as a signaling molecule in mammals. We hypothesized that oleic acid (Ole) may stimulate the formation of satellite cell-derived intramuscular adipose tissue. The objective of the current study was to determine the effect of Ole on GPR43 and factors related to the adipogenic differentiation of bovine satellite cells. Bovine satellite cells were isolated from the semimembranosus of two 14-month-old crossbreed steers. The isolated muscle satellite cells were incubated in Dulbecco’s Modified Eagle’s Medium (DMEM) solution with 10% Fetal Bovine Serum. Upon reaching 80 to 90% confluence, the growth medium was replaced with differentiation medium composed of DMEM and 2% horse serum, 10μg/mL insulin, 10μg/mL hydrocortisone, 10μM ciglitizone, and 1×antibiotic-antimycotic with dose of: 0, 1, 10, 100, or 500 μM of oleic acid (Ole). Addition of Ole on BSC induced transdifferentiation of myogenic lineage into adipocyte-like cells which formed lipid droplets within cells. Use of 100 μM and 500 μM Ole doses tended to result in a greater (P &lt; 0.1) amount of mRNA gene expression of C/EBPβ compared to all other doses. This might suppress myogenic differentiation. Expression of PPARγ was not altered (P &gt; 0.1) by treatment. The addition of 100 μM and 500 μM upregulated (P &lt; 0.05) mRNA gene expression of GPR43 and 100 μM of Ole increased protein level of GPR43 (P &lt; 0.05) and phosphorylated AMPKα (P &lt; 0.05).

scholarly journals Beneficial Effect of IL-4 and SDF-1 on Myogenic Potential of Mouse and Human Adipose Tissue-Derived Stromal Cells

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1479
Author(s):  
Karolina Archacka ◽  
Joanna Bem ◽  
Edyta Brzoska ◽  
Areta M. Czerwinska ◽  
Iwona Grabowska ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Adipose Tissue ◽  
Satellite Cells ◽  
Stromal Cells ◽  
Muscle Regeneration ◽  
Myogenic Differentiation ◽  
Human Adipose Tissue ◽  
Skeletal Muscle Regeneration ◽  
Animal Cells ◽  
The Impact

Under physiological conditions skeletal muscle regeneration depends on the satellite cells. After injury these cells become activated, proliferate, and differentiate into myofibers reconstructing damaged tissue. Under pathological conditions satellite cells are not sufficient to support regeneration. For this reason, other cells are sought to be used in cell therapies, and different factors are tested as a tool to improve the regenerative potential of such cells. Many studies are conducted using animal cells, omitting the necessity to learn about human cells and compare them to animal ones. Here, we analyze and compare the impact of IL-4 and SDF-1, factors chosen by us on the basis of their ability to support myogenic differentiation and cell migration, at mouse and human adipose tissue-derived stromal cells (ADSCs). Importantly, we documented that mouse and human ADSCs differ in certain reactions to IL-4 and SDF-1. In general, the selected factors impacted transcriptome of ADSCs and improved migration and fusion ability of cells in vitro. In vivo, after transplantation into injured muscles, mouse ADSCs more eagerly participated in new myofiber formation than the human ones. However, regardless of the origin, ADSCs alleviated immune response and supported muscle reconstruction, and cytokine treatment enhanced these effects. Thus, we documented that the presence of ADSCs improves skeletal muscle regeneration and this influence could be increased by cell pretreatment with IL-4 and SDF-1.

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