Vitamin D deficiency down-regulates Notch pathway contributing to skeletal muscle atrophy in old Wistar rats

(1)

HAL Id: hal-01216756

https://hal.archives-ouvertes.fr/hal-01216756

Submitted on 12 Nov 2015

HAL is a multi-disciplinary open access

archive for the deposit and dissemination of sci-entific research documents, whether they are pub-lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers.

L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Vitamin D deficiency down-regulates Notch pathway contributing to skeletal muscle atrophy in old Wistar

rats

Carla Domingues-Costa Faria, Audrey Chanet, Jérôme Salles, Alexandre Berry, Christophe Giraudet, Véronique Patrac, Philippe Denis, Katia Bouton,

Nicolas Goncalves-Mendes, Marie-Paule Vasson, et al.

To cite this version:

Carla Domingues-Costa Faria, Audrey Chanet, Jérôme Salles, Alexandre Berry, Christophe Giraudet, et al.. Vitamin D deficiency down-regulates Notch pathway contributing to skeletal muscle atrophy in old Wistar rats. IFFC-EFLM EUROMEDLAB Paris 2015, Jun 2015, Paris, France. 2015. �hal-01216756�

(2)

0,0 0,2 0,4 0,6 0,8 1,0 1,2 1,4 Control Depleted Delta -1 mRNA fold chan ge 0 10 20 30 40 50 60 Control Depleted Plasm a 25(OH)D conc entrati on (nM ) 0 20 40 60 80 100 120 140 Control Depleted VDR protein expre ssio n (A.U.) * 0 2 4 6 8 10 12 14 Control Depleted TA ma ss/b ody weight (x10 -4 ) *

Vitamin D deficiency down-regulates Notch pathway contributing to

skeletal muscle atrophy in old Wistar rats

Domingues-Faria C.1,2,4_{, Chanet A.}2,4_{, Salles J.}2,4_{, Berry A.}2,4_{, Giraudet C.}2,4_{, Patrac V.}2,4_{, Denis P.}3,4_{, Bouton K.}2,4_{, Goncalves-Mendes N.}1_,

Vasson M.-P.1,5_{, Boirie Y.}2,6_{, Walrand S.}2,4

Clermont Université, Université d'Auvergne, UMR 1019 INRA-UdA, Unité de Nutrition Humaine, Equipe ECREIN1_{, Equipe NuTriM}2_{, Installation expérimentale de Nutrition}3_{, CRNH-Auvergne, Clermont-Ferrand.}

4_{INRA, UMR1019, UNH, CRNH Auvergne, Clermont-Ferrand.}3 _{CHU Clermont-Ferrand, Centre Jean Perrin, Unité de Nutrition, CLARA, Clermont-Ferrand.}6_{CHU Clermont-Ferrand, Service de Nutrition Clinique,}

Clermont-Ferrand

INTRODUCTION: The aged-related decrease in skeletal muscle regeneration capacity is a contributing factor to sarcopenia [1]. Muscle

regeneration depends in part on satellite cells whose reserve pool is reduced during aging [1]. The decreased activity of the Notch signalling pathway with aging likely explains the reduction in the proliferation potential of satellite cells, as this pathway is involved in self-renewal of these cells [2]. Skeletal muscle is a primary target of vitamin D action. It modulates muscle proliferation and differentiation in vitro and stimulates muscle regeneration in vivo [3-4]. The vitamin D status is positively correlated with skeletal muscle mass and function. Hypovitaminosis D frequently observed in the elderly is responsible for muscle weakness [5-6].

AIM OF THE STUDY: Evaluation of how vitamin D deficiency induces skeletal muscle atrophy in old rats through a reduction in Notch pathway activity and proliferation potential in muscle.

RESULTS:

EXPERIMENTAL PROTOCOL:

CONCLUSION : Vitamin D depletion for 9 months efficiently induced vitamin D deficiency in old rats. Vitamin D depletion induces skeletal

muscle atrophy in old rats through a reduction in the proliferative ability and in Notch pathway activity in skeletal muscle. In this context, vitamin D deficiency could further aggravate the age-related impaired capacity of muscle to regenerate.

Domingues-Faria et al. Nutrition & Metabolism 2014, 11:47

15-month-old Male Wistar Rats

(n=20)

Standard Diet

Vitamin D-depleted Diet

24-month-old Male Wistar Rats control group (n=10)

vitamin D-depleted (VDD) group (n=10)

ELISA for circulating 25(OH)D measurment, qPCR for gene expressions and western-blot for protein expressions in tibialis anterior (TA) muscle

** Plasma 25(OH)D Control VDD TA mass/ body weight 0 20 40 60 80 100 120 140 160 Control Depleted PCNA protein expre ssio n (A.U.) * PCNA Protein

(Proliferating Cell Nuclear Antigen)

Control VDD

VDR protein

Control VDD

Notch signalling pathway

* Delta-1 mRNA Control VDD 0 20 40 60 80 100 120 Control Depleted Delta -1 protein expre ssio n (A.U.) Delta-1 protein Control VDD 0,0 0,2 0,4 0,6 0,8 1,0 1,2 1,4 Control Depleted Notch -1 mRNA fold change Notch-1 mRNA Control VDD 0 50 100 150 200 250 Control Depleted (TM ) Notch -1 protein expre ssio n (A.U.) *

Cleaved Notch-1 protein

Control VDD 0,0 0,2 0,4 0,6 0,8 1,0 Control Depleted Hes1 mRNA fold change * Hes1 mRNA Control VDD

Mean ± SEM ; Statistics : T-test of Student, * p< 0,05 ou ** p< 0,01 versus control group.

After injury, Delta-like is stimulated in satellite cells and myofibers. Delta-like binds to Notch receptor which activates Notch pathway. Then two proteolytic cleavage events occur. The first involves an ADAM protease (a Disintegrin and Metalloprotease) cleaving the Notch receptor and generating a transmembrane fragment of Notch: TM_Notch. TM_{Notch is then cleaved by a} γ-secretase complex, leading to the release of the intracellular domain of Notch: Nicd_Notch. Nicd_{Notch migrates to the nucleus where it acts} as a transcription factor, stimulating the expression of target genes as Hes1. These genes are involved in the inhibition of differentiation and the maintenance of cell renewal.

Vitamin D deficiency leads to

TA mass decrease. Plasma 25(OH)D concentration is_{reduced in VDD rats.}

Vitamin D receptor (VDR)

expression is diminished in TA of VDD rats.

The expression of the

proliferation marker PCNA is reduced in TA of VDD rats.

The gene expression of Delta1 decreases in TA of VDD old rats but the protein expression is not modified.

The gene expression of Notch-1 is not

changed following vitamin D

depletion. The protein expression of cleaved Notch form is reduced in TA of VDD old rats.

The gene expression of the target of Notch-1, Hes1, is reduced in TA of VDD old rats.

Notch pathway activity is

diminished in TA muscle of VDD old rats.

[1] Barberi (2013), [2] Conboy (2003), [3] Ceglia (2009), [4] Stratos (2013), [5] Wacker (2013), [6] Holick (2006)