Contents lists available at ScienceDirect Archives of Oral Biology journal homepage: www.elsevier.com/locate/archoralbio Protein expression in submandibular glands of young rats is modified by a high-fat/high-sugar maternal diet Martine Morzel a, ⁎ , Hélène Brignot a , Franck Ménétrier a , Géraldine Lucchi b , Vincent Paillé c , Patricia Parnet c , Sophie Nicklaus a , Marie-Chantal Canivenc-Lavier a a Centre des Sciences du Goût et de l'Alimentation, AgroSup Dijon, CNRS, INRA, Université de Bourgogne Franche-Comté, F-21000, Dijon, France b CLIPP (Clinical Innovation Proteomic Platform), Pôle de Recherche Université de Bourgogne, F- 21000, Dijon, France c Physiologie des Adaptations Nutritionnelles (PhAN), UMR1280 INRA, Université de Nantes, Institut des Maladies de l’Appareil Digestif, Nantes, France ARTICLE INFO Keywords: Western diet Salivary glands Proteome Immunohistology Annexin A5 ABSTRACT Objective: Maternal diet has consequences on many organs of the offspring, but salivary glands have received little attention despite the importance of the saliva secretory function in oral health and control of food intake. The objective of this work was therefore to document in rats the impact of maternal high-fat/high-sugar diet (Western Diet) on submandibular glands of the progeny. Design: Sprague-Dawley rat dams were fed either a Western diet or control diet during gestation and lactation and their pups were sacrificed 25 days after birth. The pups’ submandibular gland protein content was char- acterized by means of 2D-electrophoresis followed by LC–MS/MS. Data were further analyzed by Gene Ontology enrichment analysis and protein-protein interactions mapping. The expression of two specific proteins was also evaluated using immunohistochemistry. Results: Combining both male and female pups (n = 18), proteome analysis revealed that proteins involved in protein quality control (e.g. heat shock proteins, proteasome sub-units) and microtubule proteins were over- expressed in Western diet conditions, which may translate intense metabolic activity. A cluster of proteins controlling oxidative stress (e.g. Glutathione peroxidases, peroxiredoxin) and enhancement of the antioxidant activity molecular function were also characteristic of maternal Western diet as well as under-expression of annexin A5. The down-regulating effect of maternal Western diet on Annexin A5 expression was significant only for males (p < 0.05). Conclusions: A maternal Western diet modifies the protein composition of the offspring’s salivary glands, which may have consequences on the salivary function. 1. Introduction Salivary glands produce and secrete saliva in the oral cavity. Because saliva has many functions including for example mineraliza- tion of teeth, lubrication of hard and soft oral surfaces or protection against pathogenic micro-organisms (Dawes et al., 2015), salivary glands are essential organs for homeostasis of the oral cavity. Saliva is also related to food intake, through its role in formation of a food bolus safe to swallow or its involvement in flavour perception (Hannig, Hannig, & Morzel, 2017; Matsuo, 2000). Diet, either considered through the intake of specific nutrients or food constituents, or through a more global view of dietary quality or diversity, has been linked to saliva composition in animal models (Cappai, Wolf, Pinna, & Kamphues, 2013; Katsukawa & Ninomiya, 1999; Lamy et al., 2010; Rodrigues et al., 2015) but also in humans (Actis, Perovic, Defago, Beccacece, & Eynard, 2005; Johansson & Birkhed, 1994; Mejean et al., 2015; Morzel et al., 2012, 2017). By comparison, the literature is less profuse to describe how salivary glands themselves are affected by diet. The size of rat salivary glands was observed to vary with the level of mastication required to ingest food (Hall & Schneyer, 1964; Johnson & Sreebny, 1982) and was impacted by the dietary content in vitamin A (Horn, Redman, & Ambudkar, 1996). Gene or protein expression has also been linked to diet. For example the expression of the Prrt1 gene (a plausible candidate gene on mouse chromosome 17 that regulates an- imal food choice) was reduced in mouse parotid glands by a macro- nutrient-selection diet vs a control chow diet (Simon et al., 2015). A high-fat diet altered the abundance of several antioxidant compounds (catalase, SOD…) in rat parotid glands (Zalewska et al., 2014) and the https://doi.org/10.1016/j.archoralbio.2018.08.021 Received 9 July 2018; Received in revised form 30 August 2018; Accepted 31 August 2018 ⁎ Corresponding author at: INRA, CSGA, 17 rue Sully, F-21000, Dijon, France. E-mail address: martine.morzel@inra.fr (M. Morzel). Archives of Oral Biology 96 (2018) 87–95 0003-9969/ © 2018 Elsevier Ltd. All rights reserved. T