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Proteomic study of lumbar spinal cord after quadricipital eccentric exercise

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Proteomic study of lumbar spinal cord after

quadricipital eccentric exercise

Zoé Lacrosse1, 2, Stéphanie Hody1, Pierre Leprince1, Jean-Louis Croisier2, 3, Bernard Rogister1, 4, 5

1GIGA-Neurosciences, University of Liège, Liège, Belgium 2Department of Motricity Sciences, University of Liège, Liège, Belgium 3Department of Physical Medicine, CHU of Liège, Liège, Belgium 4Department of Neurology, CHU, University of Liège, Liège, Belgium

5GIGA-Development, Stem Cells and Regenerative Medicine, University of Liège, Liège, Belgium.

Contact : zlacrosse@ulg.ac.be

Eccentric muscle contractions are characterized by an increase of muscle tension as it lengthens (slowering movements). Unaccustomed or intense eccentric exercise causes “Delayed-Onset Muscle Soreness” (DOMS). DOMS include muscle pain that appears 24 to 72 hours after exercise, but also stiffness, edema and muscle proteins release in plasma.

The only systematic intervention that brings a muscle protection against DOMS is to realize submaximal eccentric contractions with a progressively increased intensity. The mechanism of this protection, called the “Repeated Bout Effect” (RBE), is not understood. However, it is likely explained by cellular, mechanical and neural theories [Scand.J.Med.&Sci.Sports, 13, 88, 2003]. The objective of this study is to better understand which neural signal is released in the muscle synapse and which brings protection by RBE.

Introduction

Methodology

Results

C 57 BL ♂ modalities Exercise Treadmill

4 Eccentric Downhill running

4 Concentric Uphill running

4 Control / Acclima -tization (10’) Warm up (10’) +/- 15° 5cm/s to 20 cm/s Run (8 bouts) 5’ run + 2’ recovery +/-15° 20 cm/s Run (10 bouts) 5’ run + 2’ recovery +/-20° 20 cm/s

1. Sacrifice = 24h after race 2. Lumbar spinal cord dissection 3. Compartmental extraction (nuclear

and cytoplasmic proteins) 4. Proteomic analysis by 2D-DIGE

coupled with mass spectrometry

Nuclear proteins 24h Cytoplasmic proteins 24h Eccentric - Control 7 Eccentric - Control 0 Concentric - Control 4 Concentric - Control 0 Eccentric - Concentric 0 Eccentric - Concentric 0

Figure 1: Comparison between eccentric and

control group

Figure 2: Comparison between concentric and

control group

Conclusion & Perspectives

We do not observe any cytoplasmic protein modification while in the nuclear extract, seven spots were more abundant in eccentric group and four in concentric group in comparison with control group.

• Tubulin (beta-2A chain & alpha-1B chain)

• Heat shock cognate 71kDa protein

• Dihydropyrimidinase-related protein 2 • Actin, cytoplasmic 1

The mass spectrometry of these proteins reveals that they are implicated in axoplasmic transport

.

At 24 hours, too few proteins modifications were detected in lumbar spinal cord, maybe as a consequence of a too short period between race and euthanasia. Implication of axoplasmic transport comforts our starting hypothesis that nervous system is able to protect muscle during the RBE by a synthesis and then a synaptic release of molecules modifying the muscle physiology.

1. Proteomic studies with sacrifice at 48h/72h/96h after race → identify when there are protein modifications in spinal cord

2. Transciptomic analysis for eccentric/concentric/control groups → identify RNA modifications in spinal cord 3. Identify which genes are implicated in muscle typology

Figure

Figure 1: Comparison  between eccentric and

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