Dataset Information

High-resolution analyses of the secretomes from murine C2C12 myotubes

ABSTRACT: Myokines released by skeletal muscle in response to contraction may contribute to the health-promoting effects of exercise. Previous studies with cultured rodent and human myotubes have revealed highly complex patterns of myokine secretion. However, the commonalities and differences in the secretory response of the different cell models have not been explored, limiting the interpretation of these results. In the present study, we performed a comprehensive analysis of contraction-regulated secretomes using the most commonly used skeletal muscle cell models.

INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Skeletal Muscle, Myotube

DISEASE(S): Type 2 Diabetes Mellitus

SUBMITTER: Michael Turewicz

LAB HEAD: Hadi Al-Hasani

PROVIDER: PXD059003 | Pride | 2025-04-11

REPOSITORIES: Pride

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			Action	DRS
	07072023_L_Aurora_2hDDA-15K_SecrEx_6hEPS_RP-S-Pep_ctrl_A4177_1.raw	Raw
	07072023_L_Aurora_2hDDA-15K_SecrEx_6hEPS_RP-S-Pep_ctrl_A4177_2.raw	Raw
	07072023_L_Aurora_2hDDA-15K_SecrEx_6hEPS_RP-S-Pep_ctrl_A4179_1.raw	Raw
	07072023_L_Aurora_2hDDA-15K_SecrEx_6hEPS_RP-S-Pep_ctrl_A4179_2.raw	Raw
	07072023_L_Aurora_2hDDA-15K_SecrEx_6hEPS_RP-S-Pep_ctrl_A4181_1.raw	Raw

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Project description:Steady-state RNA levels are a result of RNA synthesis and degradation. The importance of transcription-factor mediated induction or repression of mRNA synthesis is well established, but the role and mechanisms of RNA degradation are less well understood. We globally evaluated the RNA decay rates in proliferating and differentiated mouse myoblasts on whole-genome Affymetrix exon arrays, allowing for the assessment of directionality of RNA degradation and the detection of splice variant-specific differences in RNA decay rates. We found large differences in decay rates. mRNAs coding for proteins involved in signal transduction and transcriptional regulation have shortest half lives, whereas mRNAs coding for DNA replication enzymes and muscle contraction proteins are among the most stable. Many genes differentially expressed between proliferating and differentiated myoblasts demonstrate major differences in RNA decay rates. Quantitative PCR experiments confirmed the higher stability of transcripts with increased expression levels. RNA degradation has no apparent preferential directionality. RNA degradation appears to affect the ratio of different splice variants. For example, Itga7 isoforms with higher abundance in differentiated than in proliferating cells are more stable in differentiated cells, despite the sharing of a common 3’ untranslated region. Thus, where it was previously thought that the abundance of different splice isoforms was mainly controlled by tissue-specific splicing factors, we now demonstrate that isoforms may be produced at comparable levels but degraded with different efficiencies, depending on the differentiation status of the cells. Our results indicate that control of RNA degradation rates contributes significantly to the differentiation stage-dependent differences in abundance of transcripts and splice variants. Keywords: total RNA expression profiling; cultured cells We analyzed proliferating C2C12 cells and C2C12 cells differentiated into myotubes by serum starvation (8 days after induction of differentiation). We analyzed 7 time points after addition of actionmycin D (0, 10, 20, 30, 60, 150, 480 minutes). There were two biological replicates per condition.

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High-resolution analyses of the secretomes from murine C2C12 myotubes

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