Expression of microRNAs and target proteins in skeletal muscle of rats selectively bred for high and low running capacity

Samuel K. Pinto, Séverine Lamon, Erin Stephenson, Ming Kalanon, Jasmine Mikovic, Lauren G. Koch, Steven L. Britton, John A. Hawley, Donny M. Camera

Research output: Contribution to journalArticle

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Abstract

Impairments in mitochondrial function and substrate metabolism are implicated in the etiology of obesity and Type 2 diabetes. MicroRNAs (miRNAs) can degrade mRNA or repress protein translation and have been implicated in the development of such disorders. We used a contrasting rat model system of selectively bred high-(HCR) or low-(LCR) intrinsic running capacity with established differences in metabolic health to investigate the molecular mechanisms through which miRNAs regulate target proteins mediating mitochondrial function and substrate oxidation processes. Quantification of select miRNAs using the rat miFinder miRNA PCR array revealed differential expression of 15 skeletal muscles (musculus tibialis anterior) miRNAs between HCR and LCR rats (14 with higher expression in LCR; P < 0.05). Ingenuity Pathway Analysis predicted these altered miRNAs to collectively target multiple proteins implicated in mitochondrial dysfunction and energy substrate metabolism. Total protein abundance of citrate synthase (CS; miR-19 target) and voltage-dependent anion channel 1 (miR-7a target) were higher in HCR compared with LCR cohorts (~57 and ~26%, respectively; P < 0.05). A negative correlation was observed for miR-19a-3p and CS (r = 0.32, P = 0.015) protein expression. To determine whether miR-19a-3p can regulate CS in vitro, we performed luciferase reporter and transfection assays in C2C12 myotubes. MiR-19a-3p binding to the CS untranslated region did not change luciferase reporter activity; however, miR-19a-3p transfection decreased CS protein expression (~70%; P < 0.05). The differential miRNA expression targeting proteins implicated in mitochondrial dysfunction and energy substrate metabolism may contribute to the molecular basis, mediating the divergent metabolic health profiles of LCR and HCR rats.

Original languageEnglish (US)
Pages (from-to)E335-E343
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume313
Issue number3
DOIs
StatePublished - Sep 1 2017

Fingerprint

MicroRNAs
Running
Skeletal Muscle
Proteins
Luciferases
Energy Metabolism
Transfection
Voltage-Dependent Anion Channel 1
Untranslated Regions
Citrate (si)-Synthase
Metabolome
Mitochondrial Proteins
Skeletal Muscle Fibers
Health
Protein Biosynthesis
Protein Transport
Type 2 Diabetes Mellitus
Obesity
Polymerase Chain Reaction
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Physiology (medical)

Cite this

Expression of microRNAs and target proteins in skeletal muscle of rats selectively bred for high and low running capacity. / Pinto, Samuel K.; Lamon, Séverine; Stephenson, Erin; Kalanon, Ming; Mikovic, Jasmine; Koch, Lauren G.; Britton, Steven L.; Hawley, John A.; Camera, Donny M.

In: American Journal of Physiology - Endocrinology and Metabolism, Vol. 313, No. 3, 01.09.2017, p. E335-E343.

Research output: Contribution to journalArticle

Pinto, Samuel K. ; Lamon, Séverine ; Stephenson, Erin ; Kalanon, Ming ; Mikovic, Jasmine ; Koch, Lauren G. ; Britton, Steven L. ; Hawley, John A. ; Camera, Donny M. / Expression of microRNAs and target proteins in skeletal muscle of rats selectively bred for high and low running capacity. In: American Journal of Physiology - Endocrinology and Metabolism. 2017 ; Vol. 313, No. 3. pp. E335-E343.
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AU - Stephenson, Erin

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AU - Mikovic, Jasmine

AU - Koch, Lauren G.

AU - Britton, Steven L.

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AU - Camera, Donny M.

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