Abstract

Diets of dairy cows are often based on maize silage (MS), delivering lower amounts of n-3 fatty acids (FA) compared to grass silage-based diets. The fatty acid composition of the cell membrane can affect the cell function. We evaluated the effects of an MS-based diet on bovine red blood cell (RBC) membrane FA composition and dietary effects on controlled ATP release of RBC. In trial 1, German Holstein cows were fed an MS-based total mixed ration for 24 weeks. The FA composition of RBC membranes from repeatedly taken blood samples was analysed in addition to the abundance of the RBC membrane protein flotillin-1, which is involved in, for example, cell signalling. In trial 2, four rumen fistulated MS-fed cows were abomasally infused in a 4 × 4 Latin square model with three successively increasing lipid dosages (coconut oil, linseed–safflower oil mix (EFA; rich in n-3 FA), Lutalin®, providing conjugated linoleic acids (CLA) or the combination of the supplements, EFA + CLA) for six weeks, followed by a three-week washout period. In trial 2, we analysed RBC ATP release, flotillin-1, and the membrane protein abundance of pannexin-1, which is involved in ATP release as the last part of a signalling cascade. In trial 1, the total amount of n-3 FA in RBC membranes decreased and the flotillin-1 abundance increased over time. In trial 2, the RBC n-3 FA amount was higher after the six-week infusion period of EFA or EFA + CLA. Furthermore, depending on the dosage of FA, the ATP release from RBC increased. The abundance of flotillin-1 and pannexin-1 was not affected in trial 2. It is concluded that changes of the membrane FA composition influence the RBC function, leading to altered ATP release from intact bovine RBC.

Highlights

  • The incorporation of fatty acids (FA) of different chain length and saturation plays a critical role in plasma membrane structure and function [1], and membrane protein activity [2,3,4]

  • We evaluated the effects of an maize silage (MS)-based diet on bovine red blood cell (RBC) membrane FA composition and dietary effects on controlled adenosine triphosphate (ATP) release of RBC

  • In trial 2, we analysed RBC ATP release, flotillin-1, and the membrane protein abundance of pannexin-1, which is involved in ATP release as the last part of a signalling cascade

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Summary

Introduction

The incorporation of fatty acids (FA) of different chain length and saturation plays a critical role in plasma membrane structure and function [1], and membrane protein activity [2,3,4]. Red blood cells (RBC) are often used to investigate membrane lipid composition [7] and membrane protein function [8]. RBCs regulate vascular calibre and blood flow by releasing adenosine triphosphate (ATP) in response to physiological stimuli, such as local oxygen tension and mechanical deformation [10,11]. The signalling mechanisms leading to ATP release include a whole transduction cascade with pannexin 1 (PANX1), a channel forming membrane protein, as the final conduit [12,13]. Metabolic diseases like type 2 diabetes can affect the stimulated ATP release from RBC [14]

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