Differential effect of two dietary protein sources on time course response of muscle anabolic signaling pathways in normal and insulin dysregulated horses.
Abstract: The objective of the study was to characterize the temporal changes of phosphorylation patterns of mTOR signaling proteins in response to two dietary protein sources in insulin dysregulated (ID, = 8) and non-ID ( = 8) horses. Horses were individually housed and fed timothy grass hay and 2 daily concentrate meals so that protein was the first limiting nutrient and the total diet provided 120% of daily DE requirements for maintenance. On sample days, horses randomly received 0.25 g CP/kg BW of a pelleted alfalfa (AP) or commercial protein supplement (PS). Blood samples were collected before and 30, 60, 90, 120, 150, 180, 210, 240, 300, 360, 420, and 480 min post feeding and analyzed for plasma glucose, insulin and amino acid (AA) concentrations. G muscle samples were obtained before and 90, 180, and 300 min after feeding and analyzed for relative abundance of phosphorylated mTOR pathway components using western immunoblot analysis. There was no effect of protein source on postprandial glucose and insulin responses ( ≥ 0.14) but consumption of PS elicited a 2 times larger AUC for essential AA (EAA), greater peak concentrations of EAA and a shorter time to reach peak EAA concentrations compared to AP. Abundance of phosphorylated mTOR ( = 0.08) and rpS6 ( = 0.10) tended to be ~1.5-fold greater after consumption of PS at 90 min compared to AP. Dephosphorylation patterns differed between protein sources and was slower for AP compared to PS. ID horses had a 2 times greater ( = 0.009) AUC and 3 times higher postprandial peak concentrations ( < 0.0001) for insulin compared to non-ID horses after consumption of both treatment pellets, but EAA responses were similar between groups ( = 0.53). Insulin status did not affect rpS6 or mTOR phosphorylation after consumption of either protein source ( ≥ 0.35), but phosphorylated rpS6 abundance was twice as high in ID compared to non-ID horses ( = 0.007). These results suggest that the consumption of higher quality protein sources may result in greater postprandial activation of the mTOR pathway compared to equal amounts of a forage-based protein source. Moreover, ID does not impair postprandial activation of mTOR and rpS6 proteins in horses following a protein-rich meal.
Copyright © 2022 Loos, McLeod, Vanzant, Stratton, Bohannan, Coleman, van Doorn and Urschel.
Publication Date: 2022-08-01 PubMed ID: 35978710PubMed Central: PMC9376591DOI: 10.3389/fvets.2022.896220Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research examines the effect of two dietary protein sources on muscle growth signaling pathways in horses with normal insulin regulation and those with insulin dysregulation. The findings indicate that a higher quality protein source can spur more post-meal activation of the muscle growth pathway compared to an equivalent amount of a forage-based protein source.
Research Methodology
- The study aimed to understand the changes in phosphorylation patterns (a process that can activate or deactivate a molecule’s function) of muscle growth (mTOR) signaling proteins after the ingestion of two different protein sources. These changes were studied in horses that either had normal insulin regulation or insulin dysregulation.
- The horses were housed individually and received meals comprising timothy grass hay and 2 daily concentrate meals such that protein was the first limiting nutrient. The total diet given met 120% of daily energy requirements needed for maintenance.
- On sample days, horses were given either a protein pellet derived from alfalfa or a commercial protein supplement. Multiple measurements were taken before and after feeding over a 480-minute period to analyze plasma glucose, insulin and amino acid concentrations. Some muscle samples were also taken to analyze the relative abundance of activated mTOR pathway components.
Findings
- The protein source had no effect on the horses’ post-meal glucose and insulin responses. However, the commercial protein supplement led to twice the concentration of essential amino acids in the bloodstream, reached their peak faster, and had a bigger spike compared to the alfalfa pellet.
- The activation of mTOR and ribosomal protein S6 (rpS6), both involved in muscle growth and regeneration, tended to be approximately 1.5 times higher 90 minutes after consuming the commercial protein supplement compared to the alfalfa pellet.
- The study also found that the rate of phosphorylation varied between the two protein sources, with the commercial protein supplement dephosphorylating more rapidly.
- Horses with insulin dysregulation had twice the essential amino acid response and thrice the peak concentration of insulin compared to horses with normal insulin regulation, regardless of the protein source eaten. However, essential amino acid outcomes were consistent between the two groups.
- The research also indicated that insulin regulation did not affect the activation of rpS6 or mTOR following the consumption of either protein source. Notably, the abundance of the phosphorylated protein rpS6 was twice as high in insulin dysregulated horses than in those with normal insulin function.
Implications
- The results suggest that consuming higher quality protein sources may trigger greater post-meal activation of the mTOR pathway (which plays a vital role in muscle protein synthesis) compared to equal amounts of forage-based protein.
- Importantly, the research suggests that insulin dysregulation does not hinder the activation of mTOR and rpS6 proteins in horses following a protein-rich meal. This finding could potentially help inform dietary guidance for insulin dysregulated or diabetic horses.
Cite This Article
APA
Loos CMM, McLeod KR, Vanzant ES, Stratton SA, Bohannan AD, Coleman RJ, van Doorn DA, Urschel KL.
(2022).
Differential effect of two dietary protein sources on time course response of muscle anabolic signaling pathways in normal and insulin dysregulated horses.
Front Vet Sci, 9, 896220.
https://doi.org/10.3389/fvets.2022.896220 Publication
Researcher Affiliations
- Department of Animal and Food Sciences, University of Kentucky, Lexington, KY, United States.
- Department of Animal and Food Sciences, University of Kentucky, Lexington, KY, United States.
- Department of Animal and Food Sciences, University of Kentucky, Lexington, KY, United States.
- Department of Animal and Food Sciences, University of Kentucky, Lexington, KY, United States.
- Department of Animal and Food Sciences, University of Kentucky, Lexington, KY, United States.
- Department of Animal and Food Sciences, University of Kentucky, Lexington, KY, United States.
- Equivado Consultancy B.V., Utrecht, Netherlands.
- Department of Animal and Food Sciences, University of Kentucky, Lexington, KY, United States.
Conflict of Interest Statement
DD was hired as a consultant by the funder of this project (Versele-Laga, Deinze, Belgium). He also receives a portion of the profits from Cavalor VitAmino. DD is employed by Equivado Consultancy B.V. The funding company did not participate in the analysis or the decision to publish. All authors declare that they had full autonomy and independency in the research and publishing of this work. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
References
This article includes 59 references
- Hodson N, Philp A. The Importance of mTOR Trafficking for Human Skeletal Muscle Translational Control.. Exerc Sport Sci Rev 2019 Jan;47(1):46-53.
- Yoon MS. mTOR as a Key Regulator in Maintaining Skeletal Muscle Mass.. Front Physiol 2017;8:788.
- Wilson FA, Suryawan A, Orellana RA, Kimball SR, Gazzaneo MC, Nguyen HV, Fiorotto ML, Davis TA. Feeding rapidly stimulates protein synthesis in skeletal muscle of neonatal pigs by enhancing translation initiation.. J Nutr 2009 Oct;139(10):1873-80.
- Dickinson JM, Fry CS, Drummond MJ, Gundermann DM, Walker DK, Glynn EL, Timmerman KL, Dhanani S, Volpi E, Rasmussen BB. Mammalian target of rapamycin complex 1 activation is required for the stimulation of human skeletal muscle protein synthesis by essential amino acids.. J Nutr 2011 May;141(5):856-62.
- Hara K, Yonezawa K, Weng QP, Kozlowski MT, Belham C, Avruch J. Amino acid sufficiency and mTOR regulate p70 S6 kinase and eIF-4E BP1 through a common effector mechanism.. J Biol Chem 1998 Jun 5;273(23):14484-94.
- Atherton PJ, Smith K, Etheridge T, Rankin D, Rennie MJ. Distinct anabolic signalling responses to amino acids in C2C12 skeletal muscle cells.. Amino Acids 2010 May;38(5):1533-9.
- Tang JE, Moore DR, Kujbida GW, Tarnopolsky MA, Phillips SM. Ingestion of whey hydrolysate, casein, or soy protein isolate: effects on mixed muscle protein synthesis at rest and following resistance exercise in young men.. J Appl Physiol (1985) 2009 Sep;107(3):987-92.
- Norton LE, Layman DK, Bunpo P, Anthony TG, Brana DV, Garlick PJ. The leucine content of a complete meal directs peak activation but not duration of skeletal muscle protein synthesis and mammalian target of rapamycin signaling in rats.. J Nutr 2009 Jun;139(6):1103-9.
- Norton LE, Wilson GJ, Layman DK, Moulton CJ, Garlick PJ. Leucine content of dietary proteins is a determinant of postprandial skeletal muscle protein synthesis in adult rats.. Nutr Metab (Lond) 2012 Jul 20;9(1):67.
- Graham-Thiers PM, Bowen LK. Effect of protein source on nitrogen balance and plasma amino acids in exercising horses.. J Anim Sci 2011 Mar;89(3):729-35.
- Loos CMM, McLeod KR, Stratton SC, van Doorn DA, Kalmar ID, Vanzant ES, Urschel KL. Pathways regulating equine skeletal muscle protein synthesis respond in a dose-dependent manner to graded levels of protein intake.. J Anim Sci 2020 Sep 1;98(9).
- Avruch J, Hara K, Lin Y, Liu M, Long X, Ortiz-Vega S, Yonezawa K. Insulin and amino-acid regulation of mTOR signaling and kinase activity through the Rheb GTPase.. Oncogene 2006 Oct 16;25(48):6361-72.
- Urschel KL, Escobar J, McCutcheon LJ, Geor RJ. Insulin infusion stimulates whole-body protein synthesis and activates the upstream and downstream effectors of mechanistic target of rapamycin signaling in the gluteus medius muscle of mature horses.. Domest Anim Endocrinol 2014 Apr;47:92-100.
- Warnken T, Brehm R, Feige K, Huber K. Insulin signaling in various equine tissues under basal conditions and acute stimulation by intravenously injected insulin.. Domest Anim Endocrinol 2017 Oct;61:17-26.
- Waller AP, Burns TA, Mudge MC, Belknap JK, Lacombe VA. Insulin resistance selectively alters cell-surface glucose transporters but not their total protein expression in equine skeletal muscle.. J Vet Intern Med 2011 Mar-Apr;25(2):315-21.
- Tiley HA, Geor RJ, McCutcheon LJ. Effects of dexamethasone administration on insulin resistance and components of insulin signaling and glucose metabolism in equine skeletal muscle.. Am J Vet Res 2008 Jan;69(1):51-8.
- Schuver A, Frank N, Chameroy KA, Elliott SB. Assessment of insulin and glucose dynamics by using an oral sugar test in horses. J Equine Vet Sci. (2014) 34:465u201370. 10.1016/j.jevs.2013.09.006
- Equine Endocrinology Group,. Recommendations for the Diagnosis Treatment of Equine Metabolic Syndrome (2020). Available online at: https://sites.tufts.edu/equineendogroup/files/2020/09/200592_EMS_Recommendations_Bro-FINAL.pdf (accessed March 8, 2022).
- Equine Endocrinology Group,. Recommendations for the Diagnosis Treatment of Pituitary Pars Intermedia Dysfunction (2021). Available online at: https://sites.tufts.edu/equineendogroup/files/2021/12/2021-PPID-Recommendations-V11-wo-insert.pdf (accessed March 8, 2022).
- National Research Council . Nutrient Requirements of Horses: Sixth Revised Edition. Washington, DC: The National Academies Press; (2007).
- Henneke DR, Potter GD, Kreider JL, Yeates BF. Relationship between condition score, physical measurements and body fat percentage in mares.. Equine Vet J 1983 Oct;15(4):371-2.
- Urschel KL, Escobar J, McCutcheon LJ, Geor RJ. Effect of feeding a high-protein diet following an 18-hour period of feed withholding on mammalian target of rapamycin-dependent signaling in skeletal muscle of mature horses.. Am J Vet Res 2011 Feb;72(2):248-55.
- Loos CMM, Dorsch SC, Elzinga SE, Brewster-Barnes T, Vanzant ES, Adams AA, Urschel KL. A high protein meal affects plasma insulin concentrations and amino acid metabolism in horses with equine metabolic syndrome.. Vet J 2019 Sep;251:105341.
- Dangin M, Guillet C, Garcia-Rodenas C, Gachon P, Bouteloup-Demange C, Reiffers-Magnani K, Fauquant J, Ballu00e8vre O, Beaufru00e8re B. The rate of protein digestion affects protein gain differently during aging in humans.. J Physiol 2003 Jun 1;549(Pt 2):635-44.
- Gorissen SHM, Trommelen J, Kouw IWK, Holwerda AM, Pennings B, Groen BBL, Wall BT, Churchward-Venne TA, Horstman AMH, Koopman R, Burd NA, Fuchs CJ, Dirks ML, Res PT, Senden JMG, Steijns JMJM, de Groot LCPGM, Verdijk LB, van Loon LJC. Protein Type, Protein Dose, and Age Modulate Dietary Protein Digestion and Phenylalanine Absorption Kinetics and Plasma Phenylalanine Availability in Humans.. J Nutr 2020 Aug 1;150(8):2041-2050.
- Drummond MJ, Glynn EL, Fry CS, Timmerman KL, Volpi E, Rasmussen BB. An increase in essential amino acid availability upregulates amino acid transporter expression in human skeletal muscle.. Am J Physiol Endocrinol Metab 2010 May;298(5):E1011-8.
- Boirie Y, Dangin M, Gachon P, Vasson MP, Maubois JL, Beaufru00e8re B. Slow and fast dietary proteins differently modulate postprandial protein accretion.. Proc Natl Acad Sci U S A 1997 Dec 23;94(26):14930-5.
- Witard OC, Jackman SR, Breen L, Smith K, Selby A, Tipton KD. Myofibrillar muscle protein synthesis rates subsequent to a meal in response to increasing doses of whey protein at rest and after resistance exercise.. Am J Clin Nutr 2014 Jan;99(1):86-95.
- D'Souza RF, Marworth JF, Figueiredo VC, Della Gatta PA, Petersen AC, Mitchell CJ, Cameron-Smith D. Dose-dependent increases in p70S6K phosphorylation and intramuscular branched-chain amino acids in older men following resistance exercise and protein intake.. Physiol Rep 2014 Aug 1;2(8).
- Tang JE, Phillips SM. Maximizing muscle protein anabolism: the role of protein quality.. Curr Opin Clin Nutr Metab Care 2009 Jan;12(1):66-71.
- Gibbs PG, Potter GD, Schelling GT, Kreider JL, Boyd CL. Digestion of hay protein in different segments of the equine digestive tract.. J Anim Sci 1988 Feb;66(2):400-6.
- Farley E, Potter G, Gibbs P, Schumacher J, Murray-Gerzik M. Digestion of soybean meal proteinin the equine small and large intestine at various levels of intake. J Equine Vet Sci. (1995) 15:391u20137. 10.1016/S0737-0806(07)80483-7
- Escobar J, Frank JW, Suryawan A, Nguyen HV, Davis TA. Amino acid availability and age affect the leucine stimulation of protein synthesis and eIF4F formation in muscle.. Am J Physiol Endocrinol Metab 2007 Dec;293(6):E1615-21.
- DeBoer ML, Martinson KM, Pampusch MS, Hansen AM, Wells SM, Ward C, Hathaway M. Cultured equine satellite cells as a model system to assess leucine stimulated protein synthesis in horse muscle.. J Anim Sci 2018 Feb 15;96(1):143-153.
- Anthony JC, Lang CH, Crozier SJ, Anthony TG, MacLean DA, Kimball SR, Jefferson LS. Contribution of insulin to the translational control of protein synthesis in skeletal muscle by leucine.. Am J Physiol Endocrinol Metab 2002 May;282(5):E1092-101.
- Floyd JC Jr, Fajans SS, Conn JW, Knopf RF, Rull J. Stimulation of insulin secretion by amino acids.. J Clin Invest 1966 Sep;45(9):1487-502.
- Weijzen MEG, van Gassel RJJ, Kouw IWK, Trommelen J, Gorissen SHM, van Kranenburg J, Goessens JPB, van de Poll MCG, Verdijk LB, van Loon LJC. Ingestion of Free Amino Acids Compared with an Equivalent Amount of Intact Protein Results in More Rapid Amino Acid Absorption and Greater Postprandial Plasma Amino Acid Availability Without Affecting Muscle Protein Synthesis Rates in Young Adults in a Double-Blind Randomized Trial.. J Nutr 2022 Jan 11;152(1):59-67.
- Urschel KL, Geor RJ, Waterfall HL, Shoveller AK, McCutcheon LJ. Effects of leucine or whey protein addition to an oral glucose solution on serum insulin, plasma glucose and plasma amino acid responses in horses at rest and following exercise.. Equine Vet J Suppl 2010 Nov;(38):347-54.
- Ringmark S, Jansson A. Insulin response to feeding forage with varying crude protein and amino acid content in horses at rest and after exercise. Comp Exerc Physiol. (2013) 9:209u201317. 10.3920/CEP13014
- DeBoer ML, Hathaway MR, Kuhle KJ, Weber PSD, Reiter AS, Sheaffer CC, Wells MS, Martinson KL. Glucose and Insulin Response of Horses Grazing Alfalfa, Perennial Cool-Season Grass, and Teff Across Seasons.. J Equine Vet Sci 2018 Sep;68:33-38.
- D'Hulst G, Masschelein E, De Bock K. Dampened Muscle mTORC1 Response Following Ingestion of High-Quality Plant-Based Protein and Insect Protein Compared to Whey.. Nutrients 2021 Apr 21;13(5).
- Anthony TG, McDaniel BJ, Knoll P, Bunpo P, Paul GL, McNurlan MA. Feeding meals containing soy or whey protein after exercise stimulates protein synthesis and translation initiation in the skeletal muscle of male rats.. J Nutr 2007 Feb;137(2):357-62.
- West DW, Burd NA, Coffey VG, Baker SK, Burke LM, Hawley JA, Moore DR, Stellingwerff T, Phillips SM. Rapid aminoacidemia enhances myofibrillar protein synthesis and anabolic intramuscular signaling responses after resistance exercise.. Am J Clin Nutr 2011 Sep;94(3):795-803.
- Anthony JC, Yoshizawa F, Anthony TG, Vary TC, Jefferson LS, Kimball SR. Leucine stimulates translation initiation in skeletal muscle of postabsorptive rats via a rapamycin-sensitive pathway.. J Nutr 2000 Oct;130(10):2413-9.
- Escobar J, Frank JW, Suryawan A, Nguyen HV, Kimball SR, Jefferson LS, Davis TA. Physiological rise in plasma leucine stimulates muscle protein synthesis in neonatal pigs by enhancing translation initiation factor activation.. Am J Physiol Endocrinol Metab 2005 May;288(5):E914-21.
- Murgas Torrazza R, Suryawan A, Gazzaneo MC, Orellana RA, Frank JW, Nguyen HV, Fiorotto ML, El-Kadi S, Davis TA. Leucine supplementation of a low-protein meal increases skeletal muscle and visceral tissue protein synthesis in neonatal pigs by stimulating mTOR-dependent translation initiation.. J Nutr 2010 Dec;140(12):2145-52.
- Yoshizawa F, Hirayama S, Sekizawa H, Nagasawa T, Sugahara K. Oral administration of leucine stimulates phosphorylation of 4E-bP1 and S6K 1 in skeletal muscle but not in liver of diabetic rats.. J Nutr Sci Vitaminol (Tokyo) 2002 Feb;48(1):59-64.
- Atherton PJ, Etheridge T, Watt PW, Wilkinson D, Selby A, Rankin D, Smith K, Rennie MJ. Muscle full effect after oral protein: time-dependent concordance and discordance between human muscle protein synthesis and mTORC1 signaling.. Am J Clin Nutr 2010 Nov;92(5):1080-8.
- van Vliet S, Beals JW, Holwerda AM, Emmons RS, Goessens JP, Paluska SA, De Lisio M, van Loon LJC, Burd NA. Time-dependent regulation of postprandial muscle protein synthesis rates after milk protein ingestion in young men.. J Appl Physiol (1985) 2019 Dec 1;127(6):1792-1801.
- Gazzaneo MC, Orellana RA, Suryawan A, Tuckow AP, Kimball SR, Wilson FA, Nguyen HV, Torrazza RM, Fiorotto ML, Davis TA. Differential regulation of protein synthesis and mTOR signaling in skeletal muscle and visceral tissues of neonatal pigs after a meal.. Pediatr Res 2011 Sep;70(3):253-60.
- Liu J, Stevens PD, Li X, Schmidt MD, Gao T. PHLPP-mediated dephosphorylation of S6K1 inhibits protein translation and cell growth.. Mol Cell Biol 2011 Dec;31(24):4917-27.
- Zhang X, Ma D, Caruso M, Lewis M, Qi Y, Yi Z. Quantitative phosphoproteomics reveals novel phosphorylation events in insulin signaling regulated by protein phosphatase 1 regulatory subunit 12A.. J Proteomics 2014 Sep 23;109:63-75.
- Tremblay F, Krebs M, Dombrowski L, Brehm A, Bernroider E, Roth E, Nowotny P, Waldhu00e4usl W, Marette A, Roden M. Overactivation of S6 kinase 1 as a cause of human insulin resistance during increased amino acid availability.. Diabetes 2005 Sep;54(9):2674-84.
- de Laat MA, McGree JM, Sillence MN. Equine hyperinsulinemia: investigation of the enteroinsular axis during insulin dysregulation.. Am J Physiol Endocrinol Metab 2016 Jan 1;310(1):E61-72.
- Loos C, Dorsch S, Gerritsen A, Barnes T, Urschel K. Effects of short-term dexamethasone administration on glucose and insulin dynamics and muscle protein signaling in horses after the consumption of a high protein meal. J Anim Sci. (2018) 96:250u20131. 10.1093/jas/sky404.548
- Mu00f8ller N, Nair KS. Diabetes and protein metabolism.. Diabetes 2008 Jan;57(1):3-4.
- Nair KS, Garrow JS, Ford C, Mahler RF, Halliday D. Effect of poor diabetic control and obesity on whole body protein metabolism in man.. Diabetologia 1983 Nov;25(5):400-3.
- Pereira S, Marliss EB, Morais JA, Chevalier S, Gougeon R. Insulin resistance of protein metabolism in type 2 diabetes.. Diabetes 2008 Jan;57(1):56-63.
- Loos CMM, Vanzant ES, Bohannan AD, Urschel KL, Klotz JL, McLeod KR. Impact of ergot alkaloids on insulin sensitivity in normal and insulin dysregulated horses. In: Proceedings of the annual meeting of the European Society of Veterinary and Comparative Nutrition; 2021 Sept 9-11; Virtual meeting (2021).