Maternal Metabolizable Protein and Rumen-Protected Methionine Supplementation Impacts Steer Gene Expression in Muscle

Published: April 8, 2022

By: K Acton, IB Mandell, L Huber, MA Steele, and KM Wood / Department of Animal Biosciences, University of Guelph, Guelph, ON.

Summary

Maternal nutrition can influence fetal growth in utero and progeny development throughout life. To assess how maternal nutrition during late gestation impacts mRNA expression of genes related to growth, myogenesis, and adipogenesis in steers, 138 Angus cows were used in a 3x2 factorial arrangement of treatments (Exp. 1). For the last 8 wks of gestation, cows were randomly assigned to be fed at 90, 100, or 110% of metabolizable protein (MP) requirements, with(without) 9g/d of rumen-protected methionine (RPM). In addition, to assess similar outcomes in an industry applied setting, a 2nd study of 67 Angus cows had unlimited access to water and hay and received 0.75kg/hd/d of supplemental pellet supplying 12g/d of rumenprotected methionine (MET), or an identical pellet without additional methionine (CON; Exp. 2). After weaning, steer progeny (n1=51, n2=34) were assigned to pens by weight and fed a grower diet (58% corn silage/26% haylage/15% SBM) for 47d, followed by a finisher diet (78% high moisture corn/12% haylage/8% SBM) for 115 ±31.5d until slaughter. Muscle samples were collected for mRNA analysis. Data analyzed using PROC GLIMMIX in SAS, with maternal treatments as fixed effects and pen as a random effect. In Exp. 1, myogenin expression in muscle (MYOG) was greater (90%: 0.72, 100%: 0.94, 110%: 1.00; SEM=0.083, P=0.02) in steers from cows fed 110% MP requirements. Pyruvate kinase (PKM) expression (associated with lean tissue growth) was reduced (RPM: 0.81, NoRPM: 1.16; SEM=0.092, P=0.02) in steers from RPM supplemented cows. In Exp. 2, MET supplementation increased PKM expression (MET: 1.98, CON: 1.35; SEM=0.221, P=0.05) and myoblast determination protein-1 (MYOD; MET: 1.82, CON: 1.20; SEM=0.190, P=0.02) in muscle. These data suggest that maternal nutrition in late gestation influences offspring muscle development throughout life, which could influence carcass grade and muscle characteristics.

Key words: developmental programming, methionine, metabolizable protein, maternal nutrition, myogenin.

Introduction

Maternal nutrition during gestation and early life events impact offspring development throughout life, in a phenomenon known as developmental programming. The Barker hypothesis explains that undernutrition during specific developmental periods in pregnancy can create a scenario of pre- and post-partum metabolic mismatch (Hales and Barker, 2001). Metabolic mismatch occurs when offspring of undernourished cows are ‘programed’ to expect limited nutritional resources pre-partum, but postnatally consume diets that meet or exceeded nutritional requirements, which can influence fat and muscle development (Godfrey et al., 2007). In late gestation, fetal growth exponentially increases (Prior and Laster, 1979), additionally, offspring muscle fibres are maturing at this time, and intramuscular adipocytes (which accumulate fat throughout life and leads to marbling) begin to form (Du et al., 2010). Therefore, dietary manipulations specifically during late gestation may have a lasting impact on offspring development and meat quality. Previous studies show that maternal crude protein supplementation in late gestation influences steer offspring performance in the feedlot and carcass quality (Stalker et al., 2007; Larson et al., 2009). However, minimal research has been conducted to assess how maternal metabolizable protein (MP) alone influences steer offspring throughout life. Hare et al. (2019) observed that feeding cows 133% MP requirements did not impact gene expression (insulin-like growth factors (IGFs), insulin receptor (INSR), myogenic differentiation (MYOD), myogenin (MYOG)) in muscle from heifer calves at weaning. However, further examination is required to determine how maternal MP level influences steers in the feedlot. Methionine is an essential amino acid and is involved in many metabolic pathways in the body (Jacometo et al., 2017) and supplementation of methionine in maternal diets can alter offspring gene expression (Jacometo et al., 2017; Zhang, 2018). However, the majority of research investigating the impact of methionine on offspring have only followed dairy (Batistel et al., 2017; Alharthi et al., 2018) and beef (Clements et al., 2017; Moriel et al., 2020) calves from birth until weaning at 8 weeks and 6 months of age, respectively. Therefore, research to determine how maternal methionine supplementation impacts offspring development past weaning is warranted. The main objectives of this study were to assess how maternal metabolizable protein concentration and rumen-protected methionine supplementation in late gestation impacts beef steer offspring development in the feedlot, and carcass quality. To gain further insight into potential mechanisms involved in offspring development, the expression of genes associated with growth, muscle development, and fat accumulation were assessed.

Methods

In a research-intensive study (Exp. 1), 138 Angus cross cows were randomly assigned to one of three protein treatments, 90, 100, or 110% of metabolizable protein (MP) requirements. Half of each group was received pellets containing nine g/d of rumen-protected methionine (RPM, Smartamine^®M), or no additional pellets (NoRPM). These diets were formulated to be isocaloric through the addition of palm fat (cow performance data previously reported by Collins et al., 2019). In a second study (Exp. 2) to evaluate how maternal nutrition impacts offspring in an industry applied setting, 67 Angus cross cows received 0.75 kg/head/d of pellet supplying 12 g/d of rumen-protected methionine (MET, Smartamine®M), or a similar pellet without additional methionine (CON). Maternal diets for both studies were fed for the last eight weeks of gestation. After weaning, steer offspring (Exp. 1 n=55, Exp. 2 n=34) were managed as a group in the feedlot and fed common diets formulated to meet or exceed the requirements for growing and finishing beef steers (NASEM, 2016). Body weights were recorded every 14 days, blood samples were collected three days before slaughter, and carcasses were graded by a certified grader 24 hours after slaughter. Samples of neck muscle (Sternomandibularis muscle) were collected at slaughter for mRNA expression of genes associated with growth (IGFs, INSR and pyruvate kinase (PKM)) and the development of muscle (MYOD and MYOG) and fat (peroxisome proliferator activated receptor gamma (PPARG)) using real-time PCR (as described in Paradis et al., 2017; Hare et al., 2019). Data from the research-intensive and industry applied studies were analyzed separately using PROC GLIMMIX in SAS (University Edition, Version 9.4; SAS Institute Inc., Cary, NC) with maternal dietary treatments as the fixed effects, and steer and pen as the random effect.

Results and Discussion

Exp. 1: Steers from cows fed below or at MP requirements had higher (90%: 0.34 mmol/L, 100%: 0.39 mmol/L, 110%: 0.21 mmol/L; SEM = 0.041, P = 0.01) circulating non-esterified fatty acid (NEFA) concentrations, which may be indicative of increased fat mobilization (Webb et al., 1969). Steers from cows fed at 90% MP also had higher (90%: 15.5 mm, 100%: 14.7 mm, 110%: 11.8 mm; SEM = 1.11, P = 0.04) carcass grade fat. These results support metabolic mismatch theory, in which undernutrition during gestation followed by diets that meet requirements postnatally increases fat development (Hales and Barker, 2001; Godfrey et al., 2007). Steers from cows fed over MP requirements had increased expression of MYOG (90%: 0.72, 100%: 0.94, 110%: 1.00; SEM = 0.083, P = 0.02), which is involved in muscle development (Paradis et al., 2017). Maternal MP level did not influence any other genes associated with growth (P > 0.09), fat (P = 0.30), or muscle (P = 0.57) development. Maternal methionine supplementation reduced the expression of PKM (RPM: 0.81, NoRPM: 1.16; SEM = 0.092, P = 0.01) in steer offspring muscle.

Exp. 2: Pyruvate kinase expression in offspring from methionine supplemented cows in the industry applied study had a higher (MET: 1.98, CON: 1.35; SEM = 0.221, P = 0.05) expression of PKM and was opposite of the effect in Exp. 1. This difference could be due to maternal management practices, as cows from the industry applied study had ad libitum access to their feed and dams from Exp. 1 feed intake was controlled. Steer offspring from MET cows also had consistently heavier body weight (P ≤ 0.02); which may be reflective of increased PKM expression, which is associated with tissue growth (Hamelin et al., 2006) and muscle development (Lametsch et al., 2006). Additionally, steers from MET cows had increased (MET: 1.82, CON: 1.20; SEM = 0.190, P = 0.02) expression of MYOD which is also involved in muscle development (Paradis et al., 2017).

Conclusions

These studies show that maternal nutrition in late gestation influences beef steer offspring fat and muscle development. Feeding cows below MP requirements in late gestation, followed by diets that met requirements postnatally, influenced steer offspring fat development, which supports metabolic mismatch of the Barker hypothesis. Maternal nutrition in late gestation may prove to be a promising management opportunity to improve offspring development, carcass quality, and may lead to more AAA Canadian steaks.

Acknowledgements

We would like to thank the Elora Beef Research Centre, New Liskeard Agricultural Research Station, and University of Guelph Meat Lab staff, as well as all laboratory technicians for their assistance in this project. We would also like to thank the Natural Sciences and Engineering Research Council of Canada, Agriculture and Agri-Food Canada, and Beef Farmers of Ontario for their funding support.

Presented at the 2021 Animal Nutrition Conference of Canada. For information on the next edition, click here.

References

Alharthi, A. S., F. Batistel, M. K. Abdelmegeid, G. Lascano, C. Parys, A. Helmbrecht, E. Trevisi, and J. J. Loor. 2018. Maternal supply of methionine during late-pregnancy enhances rate of Holstein calf development in utero and postnatal growth to a greater extent than colostrum source. J. Anim. Sci. Biotechnol. 9:83. doi:10.1186/s40104-018-0298-1.

Batistel, F., A. S. Alharthi, L. Wang, C. Parys, Y.-X. Pan, F. C. Cardoso, and J. J. Loor. 2017. Placentome Nutrient Transporters and Mammalian Target of Rapamycin Signaling Proteins Are Altered by the Methionine Supply during Late Gestation in Dairy Cows and Are Associated with Newborn Birth Weight. J. Nutr. jn251876. doi:10.3945/jn.117.251876.

Clements, A. R., F. A. Ireland, T. Freitas, H. Tucker, and D. W. Shike. 2017. Effects of supplementing methionine hydroxy analog on beef cow performance, milk production, reproduction, and preweaning calf performance1. J. Anim. Sci. 95:5597–5605. doi:10.2527/jas2017.1828.

Collins, M. M., M. K. S. Lievre, K. V. J. Lawson, I. B. Mandell, A.-K. Shoveller, and K. M. Wood. 2019. Does supplemental protein and rumen-protected methionine improve performance and digestibility during late-gestation in beef cows? Available from: https://academic.oup.com/jas/article/97/Supplement_3/75/5665909

Du, M., J. Tong, J. Zhao, K. R. Underwood, M. Zhu, S. P. Ford, and P. W. Nathanielsz. 2010. Fetal programming of skeletal muscle development in ruminant animals1. J. Anim. Sci. 88:E51–E60. doi:10.2527/jas.2009-2311.

Godfrey, K. M., K. A. Lillycrop, G. C. Burdge, P. D. Gluckman, and M. A. Hanson. 2007. Epigenetic Mechanisms and the Mismatch Concept of the Developmental Origins of Health and Disease. Pediatr. Res. 61:5R-10R. doi:10.1203/pdr.0b013e318045bedb.

Hales, C. N., and D. J. P. Barker. 2001. The thrifty phenotype hypothesis. Br. Med. Bull. 60:5– 20. doi:10.1093/bmb/60.1.5.

Hamelin, M., T. Sayd, C. Chambon, J. Bouix, B. Bibe, D. Milenkovic, H. Leveziel, M. Georges, A. Clop, P. Marinova, and E. Laville. 2006. Proteomic analysis of ovine muscle hypertrophy. J Anim Sci. 84:3266–3276.

Hare, K. S., K. M. Wood, C. Fitzsimmons, and G. B. Penner. 2019. Oversupplying metabolizable protein in late gestation for beef cattle: effects on postpartum ruminal fermentation, blood metabolites, skeletal muscle catabolism, colostrum composition, milk yield and composition, and calf growth performance1. J. Anim. Sci. 97:437–455. doi:10.1093/jas/sky413.

Jacometo, C. B., Z. Zhou, D. Luchini, M. N. Corrêa, and J. J. Loor. 2017. Maternal supplementation with rumen-protected methionine increases prepartal plasma methionine concentration and alters hepatic mRNA abundance of 1-carbon, methionine, and transsulfuration pathways in neonatal Holstein calves. J. Dairy Sci. 100:3209–3219. doi:10.3168/jds.2016-11656.

Lametsch, R., L. Kristensen, M. R. Larsen, M. Therkildsen, N. Oksbjerg, and P. Ertbjerg. 2006. Changes in the muscle proteome after compensatory growth in pigs. J Anim Sci. 84:918– 924.

Larson, D. M., J. L. Martin, D. C. Adams, and R. N. Funston. 2009. Winter grazing system and supplementaion during late gestation influence performance of beef cows and steer progeny. J. Anim. Sci. 87:1147–1155. doi:10.2527/jas.2008-1323.

Moriel, P., M. Vedovatto, E. A. Palmer, R. A. Oliveira, H. M. Silva, J. Ranches, and J. M. B. Vendramini. 2020. Maternal supplementation of energy and protein, but not methionine hydroxy analogue, enhanced postnatal growth and response to vaccination in Bos indicusinfluenced beef offspring. J. Anim. Sci. skaa123. doi:10.1093/jas/skaa123.

NASEM. 2016. Nutritional Requirements of Beef Cattle. 8th ed. National Academies Press, Washington, DC.

Paradis, F., K. M. Wood, K. C. Swanson, S. P. Miller, B. W. McBride, and C. Fitzsimmons. 2017. Maternal nutrient restriction in mid-to-late gestation influences fetal mRNA expression in muscle tissues in beef cattle. BMC Genomics. 18:632. doi:10.1186/s12864-017-4051-5.

Prior, R. L., and D. B. Laster. 1979. DEVELOPMENT OF THE BOVINE FETUS l. J. Anim. Sci. 48:1546–1553.

Stalker, L. A., L. A. Ciminski, D. C. Adams, T. J. Klopfenstein, and R. T. Clark. 2007. Effects of Weaning Date and Prepartum Protein Supplementation on Cow Performance and Calf Growth. Rangel. Ecol. Manag. 60:578–587. doi:10.2111/06-082R1.1.

Webb, D. W., H. H. Head, and C. J. Wilcox. 1969. Effect of Age and Diet on Fasting Blood and Plasma Glucose Levels, Plasma Nonesterified Fatty Acid Levels, and Glucose Tolerance in Dairy Calves. J. Dairy Sci. 52:2007–2013. doi:10.3168/jds.S0022-0302(69)86887-6.

Zhang, N. 2018. Role of methionine on epigenetic modification of DNA methylation and gene expression in animals. Anim. Nutr. 4:11–16. doi:10.1016/j.aninu.2017.08.009.

Content from the event: