scholarly journals Effect of differentiation, de novo innervation, and electrical pulse stimulation on mRNA and protein expression of Na+,K+-ATPase, FXYD1, and FXYD5 in cultured human skeletal muscle cells

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247377
Author(s):  
Vid Jan ◽  
Katarina Miš ◽  
Natasa Nikolic ◽  
Klemen Dolinar ◽  
Metka Petrič ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Mrna Expression ◽  
Human Skeletal Muscle ◽  
De Novo ◽  
Muscle Cells ◽  
Electrical Pulse ◽  
Electrical Pulse Stimulation ◽  
Human Skeletal Muscle Cells ◽  
Pulse Stimulation ◽  
Low Serum

Denervation reduces the abundance of Na+,K+-ATPase (NKA) in skeletal muscle, while reinnervation increases it. Primary human skeletal muscle cells, the most widely used model to study human skeletal muscle in vitro, are usually cultured as myoblasts or myotubes without neurons and typically do not contract spontaneously, which might affect their ability to express and regulate NKA. We determined how differentiation, de novo innervation, and electrical pulse stimulation affect expression of NKA (α and β) subunits and NKA regulators FXYD1 (phospholemman) and FXYD5 (dysadherin). Differentiation of myoblasts into myotubes under low serum conditions increased expression of myogenic markers CD56 (NCAM1), desmin, myosin heavy chains, dihydropyridine receptor subunit α1S, and SERCA2 as well as NKAα2 and FXYD1, while it decreased expression of FXYD5 mRNA. Myotubes, which were innervated de novo by motor neurons in co-culture with the embryonic rat spinal cord explants, started to contract spontaneously within 7–10 days. A short-term co-culture (10–11 days) promoted mRNA expression of myokines, such as IL-6, IL-7, IL-8, and IL-15, but did not affect mRNA expression of NKA, FXYDs, or myokines, such as musclin, cathepsin B, meteorin-like protein, or SPARC. A long-term co-culture (21 days) increased the protein abundance of NKAα1, NKAα2, FXYD1, and phospho-FXYD1Ser68 without attendant changes in mRNA levels. Suppression of neuromuscular transmission with α-bungarotoxin or tubocurarine for 24 h did not alter NKA or FXYD mRNA expression. Electrical pulse stimulation (48 h) of non-innervated myotubes promoted mRNA expression of NKAβ2, NKAβ3, FXYD1, and FXYD5. In conclusion, low serum concentration promotes NKAα2 and FXYD1 expression, while de novo innervation is not essential for upregulation of NKAα2 and FXYD1 mRNA in cultured myotubes. Finally, although innervation and EPS both stimulate contractions of myotubes, they exert distinct effects on the expression of NKA and FXYDs.

scholarly journals Electrical Pulse Stimulation of Cultured Human Skeletal Muscle Cells as an In Vitro Model of Exercise

PLoS ONE ◽  
2012 ◽  
Vol 7 (3) ◽  
pp. e33203 ◽  
Author(s):  
Nataša Nikolić ◽  
Siril Skaret Bakke ◽  
Eili Tranheim Kase ◽  
Ida Rudberg ◽  
Ingeborg Flo Halle ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
In Vitro Model ◽  
Muscle Cells ◽  
Electrical Pulse Stimulation ◽  
Human Skeletal Muscle Cells ◽  
Vitro Model ◽  
Pulse Stimulation ◽  
Stimulation Of

Electrical pulse stimulation of cultured skeletal muscle cells as a model forin vitroexercise - possibilities and limitations

2016 ◽  
Vol 220 (3) ◽  
pp. 310-331 ◽  
Author(s):  
N. Nikolić ◽  
S. W. Görgens ◽  
G. H. Thoresen ◽  
V. Aas ◽  
J. Eckel ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cells ◽  
Skeletal Muscle Cells ◽  
Electrical Pulse ◽  
Electrical Pulse Stimulation ◽  
Pulse Stimulation ◽  
Stimulation Of

Improved Anabolic Signaling In Human Skeletal Muscle Myotubes Following Electrical Pulse Stimulation

2015 ◽  
Vol 47 ◽  
pp. 127
Author(s):  
Sanghee Park ◽  
Taylor Beasley ◽  
Kristen Turner ◽  
Donghai Zheng ◽  
Chuck Tanner ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Electrical Pulse ◽  
Electrical Pulse Stimulation ◽  
Pulse Stimulation

scholarly journals Overexpression of PGC-1αIncreases Fatty Acid Oxidative Capacity of Human Skeletal Muscle Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-12 ◽  
Author(s):  
Nataša Nikolić ◽  
Magdalena Rhedin ◽  
Arild C. Rustan ◽  
Len Storlien ◽  
G. Hege Thoresen ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Lipid Metabolism ◽  
Mrna Expression ◽  
Human Skeletal Muscle ◽  
Muscle Cells ◽  
Oxidative Capacity ◽  
Skeletal Muscle Cells ◽  
Expression Of Genes ◽  
Human Myotubes ◽  
Human Skeletal Muscle Cells

We investigated the effects of PGC-1α(peroxisome proliferator-activated receptorγcoactivator-1α) overexpression on the oxidative capacity of human skeletal muscle cellsex vivo. PGC-1αoverexpression increased the oxidation rate of palmitic acid and mRNA expression of genes regulating lipid metabolism, mitochondrial biogenesis, and function in human myotubes. Basal and insulin-stimulated deoxyglucose uptake were decreased, possibly due to upregulation of PDK4 mRNA. Expression of fast fiber-type gene marker (MHCIIa) was decreased. Compared to skeletal musclein vivo, PGC-1αoverexpression increased expression of several genes, which were downregulated during the process of cell isolation and culturing. In conclusion, PGC-1αoverexpression increased oxidative capacity of cultured myotubes by improving lipid metabolism, increasing expression of genes involved in regulation of mitochondrial function and biogenesis, and decreasing expression of MHCIIa. These results suggest that therapies aimed at increasing PGC-1αexpression may have utility in treatment of obesity and obesity-related diseases.

scholarly journals Temporal analysis of reciprocal miRNA-mRNA expression patterns predicts regulatory networks during differentiation in human skeletal muscle cells

2015 ◽  
Vol 47 (3) ◽  
pp. 45-57 ◽  
Author(s):  
Rasmus J. O. Sjögren ◽  
Brendan Egan ◽  
Mutsumi Katayama ◽  
Juleen R. Zierath ◽  
Anna Krook
Keyword(s):  
Skeletal Muscle ◽  
Mrna Expression ◽  
Human Skeletal Muscle ◽  
Expression Patterns ◽  
Muscle Cells ◽  
Skeletal Muscle Cells ◽  
Altered Expression ◽  
Human Skeletal Muscle Cells ◽  
Microarray Profiling ◽  
Reciprocal Expression

microRNAs (miRNAs) are short noncoding RNAs that regulate gene expression through posttranscriptional repression of target genes. miRNAs exert a fundamental level of control over many developmental processes, but their role in the differentiation and development of skeletal muscle from myogenic progenitor cells in humans remains incompletely understood. Using primary cultures established from human skeletal muscle satellite cells, we performed microarray profiling of miRNA expression during differentiation of myoblasts ( day 0) into myotubes at 48 h intervals ( day 2, 4, 6, 8, and 10). Based on a time-course analysis, we identified 44 miRNAs with altered expression [false discovery rate (FDR) < 5%, fold change > ±1.2] during differentiation, including the marked upregulation of the canonical myogenic miRNAs miR-1, miR-133a, miR-133b, and miR-206. Microarray profiling of mRNA expression at day 0, 4, and 10 identified 842 and 949 genes differentially expressed (FDR < 10%) at day 4 and 10, respectively. At day 10, 42% of altered transcripts demonstrated reciprocal expression patterns in relation to the directional change of their in silico predicted regulatory miRNAs based on analysis using Ingenuity Pathway Analysis microRNA Target Filter. Bioinformatic analysis predicted networks of regulation during differentiation including myomiRs miR-1/206 and miR-133a/b, miRNAs previously established in differentiation including miR-26 and miR-30, and novel miRNAs regulated during differentiation of human skeletal muscle cells such as miR-138-5p and miR-20a. These reciprocal expression patterns may represent new regulatory nodes in human skeletal muscle cell differentiation. This analysis serves as a reference point for future studies of human skeletal muscle differentiation and development in healthy and disease states.

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