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Poster communications

Autophagy is required for in vitro myogenesis in piglets

Abstract : Autophagy is a lysosomal degradation pathway with a role in the turnover of cell components via self-digestion. Over the past decade, it has been recognised as an essential process to maintain cellular and energy homeostasis. Nevertheless, little attention has been paid to this process in agronomical animal species. In pigs, the role of autophagy in skeletal muscle homeostasis and more specifically on the formation of multi-nucleated muscle fibres needs to be determined. Primary culture of satellite cells, the resident muscle stem cells, is an appropriate model to investigate macro autophagy (hereafter autophagy), the main autophagy process. The objective of the current study was to evaluate tools to monitor autophagy flux in this cell model and to determine the impact of autophagy on cell differentiation. Samples of longissimus muscle were collected from 4-day-old piglets. After isolation, satellite cells were plated in growth medium, allowed to proliferate up to 80% confluence and then placed in an appropriate culture medium for 6 days to differentiate into myotubes. Proliferating and differentiating cells were explored. Autophagy-related proteins (LC3-I, LC3-II, ULK-1, Beclin, p53, PI3K and p62) were detected by Western blotting in proliferating cells and at day 1, 3 and 6 of differentiation. The two major regulators of metabolism: mammalian target of rapamycin (mTOR), a central hub of nutrient signalling and cell growth, and AMP-activated protein kinase (AMPK), a major sensor for cell energy, were also detected. Then, cells were treated with an inhibitor of autophagy flux, bafilomycin A1 to confirm that autophagy was activated during the conversion of myoblasts into myotubes in our satellite cell culture model. In conclusion, we demonstrated that basal autophagy is required for porcine satellite cell differentiation in vitro. The next step will be to demonstrate that autophagosome biogenesis impairment halts cell fusion during stages of differentiation and the fusion step of myocytes.
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Poster communications
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https://hal.inrae.fr/hal-03142047
Contributor : Emilie Bernard <>
Submitted on : Monday, February 15, 2021 - 5:05:45 PM
Last modification on : Thursday, June 3, 2021 - 9:52:01 AM

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  • HAL Id : hal-03142047, version 1

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Annie Vincent, Marie-Hélène Perruchot, Isabelle Louveau, Frederic Dessauge. Autophagy is required for in vitro myogenesis in piglets. 71. Annual Meeting of the European Federation of Animal Science (EAAP), Dec 2020, Virtual meeting, Portugal. Wageningen Academic Publishers, Book of abstracts, pp.354, 2020, Book of abstracts of the 71st annual meeting of the european federation of animal science. ⟨hal-03142047⟩

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