# In vitro effects of triiodothyronine and insulin on the metabolism of muscle cells from chickens submitted or not to heat manipulation during embryogenesis

Nathalie Couroussé, Christophe Praud, Thomas Loyau, Vincent Coustham, Sophie Tesseraud, Sonia Metayer-Coustard, Anne Collin

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Nathalie Couroussé, Christophe Praud, Thomas Loyau, Vincent Coustham, Sophie Tesseraud, et al. In vitro effects of triiodothyronine and insulin on the metabolism of muscle cells from chickens submitted or not to heat manipulation during embryogenesis. Incubation and Fertility Research Group (WP6), Aug 2017, Wageningen, Netherlands. 2017, Combined meeting of the Incubation and Fertility Research Group (WP6) and of the Fundamental Physiology and Perinatal Development Group (WP12). Abstracts. hal-01607645

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Submitted on 2 Jun 2020

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Combined meeting of the Incubation and Fertility Research Group
(IFRG/WPSA Working Group 6)
and of the
Fundamental Physiology and Perinatal Development Group (WPSA
Working Group 12)

2017 Meeting – Hof van Wageningen, the Netherlands
**August 30th – September 1st**

ABSTRACTS

Abstracts of presentations are in the order as scheduled in
programme timetable; poster abstracts are at the end of the
document

In author list * designates the presenter

Meeting sponsors

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LOHMANN
TIERZUCHT

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Pas Reform
Hatchery Technologies

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HATCHTECH
PROVIDING SUPERIOR CHICK QUALITY

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# In vitro effects of triiodothyronine and insulin on the metabolism of muscle cells from chickens submitted or not to heat manipulation during embryogenesis

N. Couroussé¹*, C. Praud¹, T. Loyau¹,², V. Coustham¹, S. Tesseraud¹, S. Métayer-Coustard¹, A. Collin¹

¹URA, INRA, 37380 Nouzilly, France, ²URZ, INRA, 97170 Petit-Bourg, Guadeloupe, France

*Email presenter: anne.collin@inra.fr

Broiler chickens are particularly sensitive to heat exposure during growth, with socio-economic consequences for poultry industry. An embryonic heat acclimation technique was developed to improve chicken long-term heat tolerance (Piestun et al., 2008). This thermal manipulation (TM) consists of a rise in temperature during the incubation of eggs 12 h/d from day 7 to 16 of incubation. TM was notably shown to change the thyroid axis regulation in the long term. Chickens resistance to an acute heat challenge was improved at slaughter age. The glucose-insulin balance of birds was also modified (Loyau et al., 2013). The aim of this work was to better understand in vitro the involvement of the thyroid hormone triiodothyronine ($T_3$) and of insulin in the regulation of metabolic pathways affecting in vivo energy and protein utilization as well as muscle growth in TM *Pectoralis major* (Loyau et al., 2014). To identify $T_3$ and insulin effects in interaction to incubation condition, we measured by qRT-PCR the expression of candidate genes involved in metabolism and muscle growth, in the regulation of muscle thyroid hormone concentrations and of oxidative stress. Signaling pathways regulating protein synthesis were also studied by biochemical method. The study was carried out both at proliferative (myoblasts) and differentiation (myotubes) cell stages in cells derived from muscle *Pectoralis major* of chicks submitted to the embryonic treatment TM or not.

In our study, TM stimulated pathways that promote proteosynthesis and modified a signaling pathway controlling cellular stress (P38 mitogen activated protein kinase). The TM also affected the regulation of energy metabolism. Indeed, the expression of Peroxisome proliferator activated receptor coactivator 1α (PGC-1α) gene involved in muscle cell energy metabolism was increased in TM as compared to control myoblasts. Apart from the observed effects of incubation conditions, direct effects of hormones were observed. $T_3$ hormone had contrasting effects on the AKT/S6K1 (S6 kinase 1)/S6 (ribosomal protein) pathway, likely stimulating proteosynthesis in myotubes. It also increased the expression of deiodinase 3 promoting $T_3$ conversion to $T_2$ (diiodothyronine). Insulin activated the AKT/S6K1/S6 pathway in both cell types and stimulated the expression of genes involved in the use of energy nutrient such as Citrate Synthase, key enzyme of the Krebs cycle, and β hydroxyacyl-Coa dehydrogenase controlling fatty acid oxidation. Furthermore, insulin interacted with incubation condition to stimulate mitochondrial metabolism via greater PGC-1α expression in TM myotubes only.

In conclusion, the in vitro study of TM-derived muscle cells revealed physiological and metabolic modifications promoting protein synthesis and energy use consecutive to embryo heat acclimation treatment in birds that were maintained through cell mitosis. Our results suggest programming mechanisms controlling gene expression under the effect of environment. We speculate that epigenetic alterations could be involved.

This study was performed with the financial support of the French Agence Nationale de la Recherche, «Jeunes Chercheuses et Jeunes Chercheurs» Project, ANR-09-JCJC-0015-01, THERMOCHICK.

## References

* Loyau et al., J Anim Sci. 2013; 91(8):3674-85. Loyau et al., PLoS One. 2014; 9(9):e105339.

* Piestun et al., Poult Sci. 2008; 87(8):1516-25.

*Keywords: Embryo acclimation, heat, $T_3$ (triiodothyronine), insulin, chicken, muscle cells, in vitro.*