New insight of some extracellular matrix molecules in beef muscles. Relationships with sensory qualities Annabelle Dubost, Didier Micol, Claire Lethias, Anne Listrat To cite this version: Annabelle Dubost, Didier Micol, Claire Lethias, Anne Listrat. New insight of some extracel- lular matrix molecules in beef muscles. Relationships with sensory qualities. animal, Pub- lished by Elsevier (since 2021) / Cambridge University Press (until 2020), 2016, 10 (5), pp.1-8. 10.1017/S1751731115002396. hal-02629905 HAL Id: hal-02629905 https://hal.inrae.fr/hal-02629905 Submitted on 27 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Animal, page 1 of 8 © The Animal Consortium 2015 animal doi:10.1017/S1751731115002396 New insight of some extracellular matrix molecules in beef muscles. Relationships with sensory qualities A. Dubost1, D. Micol1, C. Lethias2 and A. Listrat1† 1Institut National de la Recherche Agronomique (INRA), UMR1213 Herbivores, F-63122 Saint-Genès-Champanelle, France; ²Institut de Biologie et Chimie des Protéines (IBCP), FRE 3310 DyHTIT, Passage du Vercors, 69367 Lyon, Cedex 07, France (Received 4 December 2014; Accepted 28 September 2015) The aim of this study was to highlight the relationships between decorin, tenascin-X and type XIV collagen, three minor molecules of extracellular matrix (ECM), with some structural parameters of connective tissue and its content in total collagen, its cross-links (CLs) and its proteoglycans (PGs). In addition, we have evaluated impact of these minor molecules on beef quality traits. The relative abundance of these molecules was evaluated by western blot analysis in Longissimus thoracis (LT) and Biceps femoris (BF) muscles from Aberdeen Angus and Blond d’Aquitaine beef breeds. Decorin and tenascin-X were more abundant in BF than in LT (1.8 v. 0.5 arbitrary units (AU), respectively, P < 0.001, and 1.0 v. 0.6 AU, P < 0.05). There was no muscle effect for collagen XIV content. Decorin and tenascin-X relative abundance were positively correlated with perimysium and endomysium areas and with collagen characteristics (total, insoluble and CLs). Decorin was negatively correlated with total PG content and positively with tenascin-X. Collagen XIV was correlated with any of parameters measured. To assess the impact of decorin, tenascin-X and collagen XIV and of their ratios to total collagen and PGs on shear force and quality traits we realized, respectively, a multiple-linear regression analysis and a Pearson’s correlation analysis. Decorin and tenascin-X relative abundance were, respectively, negatively and positively involved in juiciness. Decorin relative abundance was also negatively involved in abnormal flavour and positively in overall liking. The ratio of decorin to total collagen and PGs was negatively correlated to juiciness, together with collagen XIV ratio to total PGs. The ratios of decorin, tenascin-X and collagen XIV to total PGs were positively correlated to sensory tenderness, negatively to abnormal beef flavour and positively to overall liking. The ratio of decorin to total collagen was also negatively correlated to abnormal flavour and positively to overall liking while its ratio to total PGs was positively correlated to beef flavour and overall liking. Results of the present study highlighted for the first time the possible role of minor ECM molecules on beef quality traits. In addition, variations of meat texture and more generally of sensory qualities would depend not only to the quantity of total collagen and of its CLs, but also of components of ECM such as decorin, tenascin-X and collagen XIV and of their ratios to total collagen and PGs. Keywords: beef, muscle, connective tissue, extracellular matrix, sensory traits Implications Introduction Consumer’s perception of meat is a critical issue for the beef Extracellular matrix (ECM) is the ground substance of industry because it impacts directly on the durability and connective tissue (CT), which is structured, by perimysium profitability of the sector. For beef consumers, tenderness, that bundles muscle fibres and by endomysium that juiciness and flavour remain the most sought sensory surrounds individual skeletal muscle fibres. ECM is a dynamic qualities. However, beef qualities are subject to variations. network of molecules mainly composed of structural After meat ageing, background toughness is partially collagens but also of many other minor components. Among explained by major components of connective tissue. In this them, we can mention collagen XIV, a minor collagen, study, the assumption was that modifications in some minor decorin, a proteoglycan (PG) and tenascin-X, a non- extracellular matrix molecules of connective tissue could collagenic glycoprotein (Velleman, 2002). They interact contribute to variations of beef qualities not explained with structural collagens and each other (Font et al., 1993; to date. Weber et al., 1996; Elefteriou et al., 2001; Lethias et al., 2006) and are co-localized in perimysium (Listrat). These † E-mail: [email protected] interactions might have important biological implications on 1 Dubost, Micol, Lethias and Listrat building of extracellular networks, alter flexibility of the ECM and insoluble collagen and CL contents than BF (Dubost and, as a result, its mechanical properties (Nishiyama et al., et al., 2013a). These muscles and these breeds had been 1994). This is confirmed by in vivo studies in mice showing chosen to create a large variability within samples. that the inactivation of the decorin or tenascin-X genes leads About structural (histology), PG and western blot analysis, to a weakness of CT (Danielson et al., 1997; Mao et al., 2002) muscle samples were taken 15 to 25 mn following exsan- and by in vitro studies showing that addition of tenascin-X or guination. For histological analysis, muscle samples of collagen XIV to collagen gels modify the mechanical prop- 1.5 × 1.5 × 1 cm were taken, fixed on a piece of cork and erties of these gels (Nishiyama et al., 1994; Margaron et al., frozen in isopentane chilled in liquid nitrogen. About PG and 2010). From this, we have hypothesized that decorin, western blot analyses, muscle samples (60 to 80 g) were cut tenascin-X and collagen XIV might, in association with up into small pieces, frozen and powdered in liquid nitrogen. structural collagens, play a role on mechanical properties of The samples were stored at −80°C until analyses. CT and thereby on meat beef quality traits. Carcasses were chilled in a cold dressing room (+2°C) and Beef meat tenderness is generally attributed to structural muscle samples of collagen and CL measurements (about collagen (generally referred to as total collagen), to its 150 g) were taken up 24 h postmortem. They were cut into cross-links (CLs) and to its structural parameters. However, pieces of 1 cm cross-section, sealed under vacuum in plastic these parameters explain only a small part of changes in bags and frozen. Frozen muscle was homogenized in a tenderness. This may be due to the fact that variations of household cutter, freeze-dried for 48 h, pulverized in a texture would depend not only of the quantity of total horizontal blade mill and finally stored at +4°C in stopper collagen and of its CLs, but also of interactions between plastic flasks until analyses. About beef quality traits sensory structural collagens and some minor components present in evaluation, meat samples were aged in vacuum packs at +4°C the ECM, such as those cited above. for 14 days, packed under vacuum and stored at −20°C until In spite of hypothetical roles of ECM molecules in Warner-Bratzler shear force measurement and sensory analysis. mechanical properties and in juiciness of poultry meat (Velleman, 2002), no direct relationship between ECM Perimysium and endomysium characteristics molecules and beef sensory quality has been reported. Muscle blocks (1.5 × 1.5 × 1cm)were fixed with Tissue-Tek® The aim of this study was to analyze: (i) the muscle OCTTM (Sakura Finetek, Villeneuve d’Ascq, France) in a cryostat variations on decorin, tenascin-X and collagen XIV relative (HM 500M; Microm Microtech, Francheville, France) maintained abundances, (ii) the relationships between these three mole- at −25°C and cross-sections of 10 µm thick were prepared. The cules, perimysium and endomysium characteristics, collagen cross-sections were stained according the Picro-Sirius red amount (total and insoluble contents) and its CLs and total method previously described by Flint and Pickering (1984). PGs and (iii) to precise the effects of these ECM molecules on Stained cross-sections were then analyzed by image analysis mechanical and sensory properties of beef meat. with two home-made programs developed by our team using Visilog 6.7 Professional Software (Noesis, Gif-sur-Yvette, France): Perimysium Analyzer to study perimysium, and Material and methods Endomysium Fibre Analyzer to study endomysium, previously The study was carried out in compliance with the French described (Dubost et al., 2013a). About perimysium study, each recommendations and those of the Animal Care and Use stained muscle section (1.5 × 1.5 cm) was scanned entirely in Committee of the National Institute for Agricultural Research transmission mode using an EPSON (Levallois Perret, France) (INRA, Institut National de la Recherche Agronomique) of Expression 10000XL PRO A3 scanner (resolution of 2400 dpi, Clermont-Ferrand/Theix, France for the use of experimental corresponding to a spatial resolution of 10.58 µm/pixel). For animals including animal welfare. endomysium, stained sections were visualized in bright field mode with an Olympus BX51 microscope (Olympus, Tokyo, Beef production and muscle sampling Japan) using a 10 × objective (NA = 0.3).