Elsevier

Theriogenology

Volume 85, Issue 9, June 2016, Pages 1669-1679
Theriogenology

Research article
Effects of rumen-protected methionine and choline supplementation on the preimplantation embryo in Holstein cows

https://doi.org/10.1016/j.theriogenology.2016.01.024Get rights and content

Abstract

Our objective was to determine the effects of supplementing methionine and choline during the prepartum and postpartum periods on preimplantation embryos of Holstein cows. Multiparous cows were assigned in a randomized complete-block design into four treatments from 21 days before calving to 30 days in milk (DIM). Treatments (TRT) were MET (n = 9, fed the basal diet + rumen-protected methionine at a rate of 0.08% [w:w] of the dry matter [DM], Smartamine M), CHO (n = 8, fed the basal diet + choline 60 g/d, Reashure), MIX (n = 11, fed the basal diet + Smartamine M and 60 g/d Reashure), and CON (n = 8, no supplementation, fed the close-up and fresh cow diets). Cows were randomly reassigned to two new groups (GRP) to receive the following diets from 31 to 72 DIM; control (CNT, n = 16, fed a basal diet) and SMT (n = 20, fed the basal diet + 0.08% [w:w] of the dry matter intake as methionine). An progesterone intravaginal insert (CIDR) device was inserted in all cows after follicular aspiration (60 DIM) and superovulation began at Day 61.5 using FSH in eight decreasing doses at 12-hour intervals over a 4-day period. On Days 63 and 64, all cows received two injections of PGF2α, and CIDR was removed on Day 65. Twenty-four hours after CIDR removal, ovulation was induced with GnRH. Cows received artificial insemination at 12 hours and 24 hours after GnRH. Embryos were flushed 6.5 days after artificial insemination. Global methylation of the embryos was assessed by immunofluorescent labeling of 5-methylcytosine, whereas lipid content was assessed by staining with Nile red. Nuclear staining was used to count the total number of cells per embryo. There was no difference between TRT, GRP, or their interaction (P > 0.05) for embryo recovery, embryos recovered, embryo quality, embryo stage, or cells per embryo. Methylation of the DNA had a TRT by GRP interaction (P = 0.01). Embryos from cows in CON-CNT had greater (P = 0.04) methylation (0.87 ± 0.09 arbitrary units [AU]) than embryos from cows in MET-CNT (0.44 ± 0.07 AU). The cytoplasmic lipid content was not affected (P > 0.05) by TRT or their interaction, but lipid content was greater (P = 0.04) for SMT (7.02 ± 1.03 AU) than that in CNT (3.61 ± 1.20 AU). In conclusion, cows in MET-CNT had embryos with lower methylation, and SMT cows had a higher lipid content than CNT. Methionine supplementation seems to impact the preimplantation embryo in a way that enhances its capacity for survival because there is strong evidence that endogenous lipid reserves serve as an energy substrate.

Introduction

Studies over the last 2 decades clearly established the link between nutrition and fertility in ruminants [1], [2], [3], [4], [5]. Dietary changes can cause an immediate and rapid alteration in a range of humoral factors that can alter endocrine and metabolic signaling pathways crucial for reproductive function [6], [7]. Moreover, periconceptional nutritional environment in humans and other animals is critical for the long-term setting of postnatal phenotype [8]. Restricting the supply of B-vitamins and methionine during the periconceptional period in sheep, e.g., resulted in adverse cardiometabolic health in postnatal offspring [9]. Feeding female mice a low-protein diet during the preimplantation period of pregnancy resulted in a reduction in amino acid (AA) concentration in uterine fluid and serum and attendant changes in the AA profile of the blastocyst [10].

Strategies have been used to improve the reproductive performance of dairy cows through alteration of nutritional status [11], [12]. In other species, dietary supplementation with specific AAs (e.g., arginine, glutamine, leucine, glycine, and methionine) had beneficial effects on embryonic and fetal survival and growth through regulation of key signaling and metabolic pathways [13], [14].

Methionine is the most limiting AA in lactating cows [15], but supplementation of diets with crystalline methionine has been excluded because free methionine is quickly and almost totally degraded by the microorganisms in the rumen [15]. In contrast, supplementing rumen-protected methionine (RPM) has a positive effect on milk protein synthesis in dairy cows [16], [17], [18]. Although the role of methionine in bovine embryonic development is unknown, there is evidence that methionine availability alters the transcriptome of bovine preimplantation embryos in vivo [19].

The DNA methylation in promoters is an important mechanism for regulation of gene expression and gene silencing. However, DNA methylation in other regions may have a more complex role in regulation of transcription [20], [21], [22]. Methylation of the DNA depends on the availability of methyl donors supplied by AAs such as methionine and by compounds of one-carbon metabolic pathways such as choline [21]. Increased methionine bioavailability is likely to increase the entry of methionine into the one-carbon metabolism cycle where it is initially converted into S-adenosylmethionine, the major biological methyl donor [23].

Choline is a major component of phospholipids, and sphingomyelin, a component of acetylcholine that participates directly in neurotransmission [24], affects membrane integrity and alters methylation pathways [25], [26]. Early studies evaluating the effect of dietary choline on milk yield and duodenal flow indicated its rapid and extensive rumen degradation before absorption in duodenum [27], [28]. Subsequently, numerous studies have evaluated the effects of feeding rumen-protected choline (RPC) on reproduction and health of dairy cows [29], [30].

Nonruminants fed diets deficient in methyl donors (e.g., choline and methionine) have hypomethylated DNA [31], [32]. These changes occur not only in global methylation [33] but also in the methylation of specific genes [34]. However, effects of methionine in preimplantation embryos are still controversial. Bonilla et al. [35] suggested that extracellular methionine is not required for DNA methylation in the cultured blastocyst. Nevertheless, gene expression changes caused by alteration of DNA methylation (i.e., absence of the methylase genes) can result in embryo death or developmental defects in preimplantation embryos [36].

The hypothesis of the present study was that dietary supplementation with RPM and RPC, or both, increases DNA methylation in preimplantation embryos in dairy cows and is beneficial to embryonic development. The objective of this study was to determine the effects of methionine and choline on DNA methylation and lipid accumulation in preimplantation embryos of Holstein cows.

Section snippets

Materials and methods

The Institutional Animal Care and Use Committee from the University of Illinois (Urbana-Champaign, IL, USA) approved all procedures performed in this experiment.

Results

The ingredient composition of the diets fed to cows is detailed in Table 1 and the analyzed chemical composition is shown in Table 2. Body weight and DMI were not affected by treatment (P > 0.17) at any time in the experiment. Values for BW were as follows: week: −3 to 0 = 770.6 kg (range: 765.6–786.2), week 1 to 4 = 675.8 kg (range: 660.5–702.5 kg), and week 5 to 10 = 640.3 kg (range: 612.7–655.7). Values for DMI were: −3 to 0 = 13.7 kg/d (range: 12.7–14.7), week 1 to 4 = 18.2 kg/d (range:

Discussion

The aim of this study was to determine the effects of RPM and RPC on development, DNA methylation, and lipid accumulation in preimplantation embryos of Holstein cows. We postulated that methionine and choline supplementation would impact the preimplantation embryo in a way that enhanced its capacity for survival so that reproductive function would be enhanced. However, neither treatment affected embryonic survival or cell number; choline also had no effect on DNA methylation or lipid

Acknowledgments

This project was supported in part by a grant from Adisseo USA. Special appreciation is extended to the staff of the University of Illinois Dairy Research Unit; Dr. Glaucio Lopes and Accelerated Genetics for providing the bull sire doses; and Dr. Richard Wallace and Zoetis for providing CIDR, Factrel, and Lutalyse used in this project.

References (60)

  • A.P. Bird et al.

    Methylation-induced repression–belts, braces, and chromatin

    Cell

    (1999)
  • M.V. Martinov et al.

    The logic of the hepatic methionine metabolic cycle

    Biochim Biophys Acta

    (2010)
  • S.H. Zeisel

    Choline: essential for brain development and function

    Adv Pediatr

    (1997)
  • M.D. Niculescu et al.

    Diet, methyl donors and DNA methylation: interactions between dietary folate, methionine and choline

    J Nutr

    (2002)
  • K.B. Atkins et al.

    Dietary choline effects on milk yield and duodenal choline flow in dairy cattle

    J Dairy Sci

    (1988)
  • B.K. Sharma et al.

    In vitro degradation of choline from selected feedstuffs and choline supplements

    J Dairy Sci

    (1989)
  • F.S. Lima et al.

    Effects of feeding rumen-protected choline on incidence of diseases and reproduction of dairy cows

    Vet J

    (2012)
  • D. Bauchart

    Lipid absorption and transport in ruminants

    J Dairy Sci

    (1993)
  • D. Rieger

    Relationships between energy metabolism and development of early mammalian embryos

    Theriogenology

    (1992)
  • A.S. Greenberg et al.

    Perilipin, a major hormonally regulated adipocyte-specific phosphoprotein associated with the periphery of lipid storage droplets

    J Biol Chem

    (1991)
  • J.C. Mathers et al.

    Induction of epigenetic alterations by dietary and other environmental factors

    Adv Genet

    (2010)
  • W.D. Rees et al.

    Maternal protein deficiency causes hypermethylation of DNA in the livers of rat fetuses

    J Nutr

    (2000)
  • M.A. Caudill et al.

    Intracellular S-adenosylhomocysteine concentrations predict global DNA hypomethylation in tissues of methyl-deficient cystathionine beta-synthase heterozygous mice

    J Nutr

    (2001)
  • S.F. Choumenkovitch et al.

    In the cystathionine beta-synthase knockout mouse, elevations in total plasma homocysteine increase tissue S-adenosylhomocysteine, but responses of S-adenosylmethionine and DNA methylation are tissue specific

    J Nutr

    (2002)
  • R.R. Grummer et al.

    Management of dry and transition cows to improve energy balance and reproduction

    J Reprod Dev

    (2010)
  • J.E. Santos et al.

    Applying nutrition and physiology to improve reproduction in dairy cattle

    Soc Reprod Fertil Suppl

    (2010)
  • T.P. Fleming et al.

    Embryos, DOHaD and David Barker

    J Dev Orig Health Dis

    (2015)
  • K.D. Sinclair et al.

    DNA methylation, insulin resistance, and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status

    Proc Natl Acad Sci U S A

    (2007)
  • J.J. Eckert et al.

    Metabolic induction and early responses of mouse blastocyst developmental programming following maternal low protein diet affecting life-long health

    PLoS One

    (2012)
  • H. DelCurto et al.

    Nutrition and reproduction: links to epigenetics and metabolic syndrome in offspring

    Curr Opin Clin Nutr Metab Care

    (2013)
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