L-Carnitine sustainably affects bioenergetic profile of bovine blastocysts and transcriptome profile of elongation-stage embryos

in Reproduction
Authors:
Eva Held-Hoelker Institute of Animal Sciences, Animal Breeding and Husbandry group, Rheinische Friedrich-Wilhelms-Universitat Bonn, Endenicher Allee, Bonn, Germany
Department of Animal Science, Biotechnology and Reproduction of Farm Animals, Georg-August-University Goettingen, Burckhardtweg, Göttingen, Germany.

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https://orcid.org/0009-0006-4732-9550
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Jessica Kurzella Institute of Animal Sciences, Animal Breeding and Husbandry group, Rheinische Friedrich-Wilhelms-Universitat Bonn, Endenicher Allee, Bonn, Germany

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Dessie Salilew-Wondim Institute of Animal Sciences, Animal Breeding and Husbandry group, Rheinische Friedrich-Wilhelms-Universitat Bonn, Endenicher Allee, Bonn, Germany
Department of Animal Science, Biotechnology and Reproduction of Farm Animals, Georg-August-University Goettingen, Burckhardtweg, Göttingen, Germany.

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Franca Rings Institute of Animal Sciences, Animal Breeding and Husbandry group, Rheinische Friedrich-Wilhelms-Universitat Bonn, Endenicher Allee, Bonn, Germany

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Dawit Tesfaye Department of Biomedical Sciences, Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, USA

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Ernst Tholen Institute of Animal Sciences, Animal Breeding and Husbandry group, Rheinische Friedrich-Wilhelms-Universitat Bonn, Endenicher Allee, Bonn, Germany

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Christine Grosse-Brinkhaus Institute of Animal Sciences, Animal Breeding and Husbandry group, Rheinische Friedrich-Wilhelms-Universitat Bonn, Endenicher Allee, Bonn, Germany

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Michael Hoelker Department of Animal Science, Biotechnology and Reproduction of Farm Animals, Georg-August-University Goettingen, Burckhardtweg, Göttingen, Germany.

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Correspondence should be addressed to E Held-Hoelker; Email: ehel@itw.uni-bonn.de
DOI:
https://doi.org/10.1530/REP-24-0004
Volume/Issue:
Volume 168: Issue 3
Article ID:
e240004
Article Type:
Research Article
Online Publication Date:
02 Aug 2024
Copyright:
© the author(s) 2024
Restricted access
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In brief

In the present study the sustainable effect of L-carnitine during the culture period on the post-transfer development was investigated. Taken together, we uncovered direct effects of L-carnitine on the bioenergetic profile of day 7 blastocysts along with sustainable effects on mtDNA copy numbers and transcriptome profile of bovine day 14 embryos.

Abstract

L-Carnitine (LC) is known to play key roles in lipid metabolism and antioxidative activity, implicating enhanced cryotolerance of bovine blastocysts. However, sustainability of LC supplementation during culture period on preimplantation development beyond the blastocyst stage has not been investigated so far. Therefore, all embryos were cultured under fatty acid-free conditions, one group with LC (LC embryos) and the control group without LC (control) supplementation. Transfer to recipients was conducted on day 6. Elongation-stage embryos were recovered on day 14; metrics of embryo recollection, developmental rates as regards early elongation-stage as well as mean embryo length did not differ between the groups. Gene expression analyses via NGS revealed 341 genes to be differentially regulated between elongation-stage embryos derived from LC supplementation compared to controls. These played mainly a role in molecular functions and biological processes like oxidoreductase activity, ATP-dependent activity, cellular stress, and respiration. Pathways like oxidative phosphorylation and thermogenesis, extracellular matrix receptor signaling, PI3K-Akt, and focal adhesion were affected by differentially regulated genes. Moreover, all DEGs located on the mitochondria were significantly downregulated in LC embryos, being in line with lower mitochondrial copy number and mtDNA integrity compared to the control group. Finally, we uncovered alterations of the bioenergetic profile on day 7 as a consequence of LC supplementation for the first time, revealing significantly higher oxygen consumption rates, ATP linked respiration and spare capacity for LC embryos. In summary, we uncovered direct effects of LC supplementation during the culture period on the bioenergetic profile along with sustainable effects on mtDNA copy numbers and transcriptome profile of bovine day 14 embryos.

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Print ISSN:
1470-1626
Online ISSN:
1741-7899

  • Abe H, Yamashita S, Satoh T & & Hoshi H 2002 Accumulation of cytoplasmic lipid droplets in bovine embryos and cryotolerance of embryos developed in different culture systems using serum-free or serum-containing media. Molecular Reproduction and Development 61 57–66. (https://doi.org/10.1002/mrd.1131)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Ali AA, Bilodeau JF & & Sirard MA 2003 Antioxidant requirements for bovine oocytes varies during in vitro maturation, fertilization and development. Theriogenology 59 939–949. (https://doi.org/10.1016/s0093-691x(0201125-1)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Arduini A, Tyurin V, Tyuruna Y, Arrigoni-Martelli E, Molajoni F, Dottori S & & Federici G 1992 Acyl-trafficking in membrane phospholipid fatty acid turnover: the transfer of fatty acid from the acyl-L-carnitine pool to membrane phospholipids in intact human erythrocytes. Biochemical and Biophysical Research Communications 187 353–358. (https://doi.org/10.1016/s0006-291x(0581500-1)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Berg DK, van Leeuwen J, Beaumont S, Berg M & & Pfeffer PL 2010 Embryo loss in cattle between days 7 and 16 of pregnancy. Theriogenology 73 250–260. (https://doi.org/10.1016/j.theriogenology.2009.09.005)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Brooks K, Burns G & & Spencer TE 2014 Conceptus elongation in ruminants: roles of progesterone, prostaglandin, interferon tau and cortisol. Journal of Animal Science and Biotechnology 5 53. (https://doi.org/10.1186/2049-1891-5-53)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Carrillo-González DF & & Maldonado-Estrada JG 2020 L-carnitine supplementation in culture media improves the pregnancy rate of in vitro produced embryos with sexed semen from Bos taurus indicus cows. Tropical Animal Health and Production 52 2559–2565. (https://doi.org/10.1007/s11250-020-02281-y)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Diez-Juan A, Rubio C, Marin C, Martinez S, Al-Asmar N, Riboldi M, Díaz-Gimeno P, Valbuena D & & Simón C 2015 Mitochondrial DNA content as a viability score in human euploid embryos: less is better. Fertility and Sterility 104 534–41.e1. (https://doi.org/10.1016/j.fertnstert.2015.05.022)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Ferré LB, Kjelland ME, Taiyeb AM, Campos-Chillon F & & Ross PJ 2020 Recent progress in bovine in vitro-derived embryo cryotolerance: impact of in vitro culture systems, advances in cryopreservation and future considerations. Reproduction in Domestic Animals = Zuchthygiene 55 659–676. (https://doi.org/10.1111/rda.13667)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Fragouli E, Spath K, Alfarawati S, Kaper F, Craig A, Michel C-E, Kokocinski F, Cohen J, Munne S & & Wells D 2015 Altered levels of mitochondrial DNA are associated with female age, aneuploidy, and provide an independent measure of embryonic implantation potential. PLoS Genetics 11 e1005241. (https://doi.org/10.1371/journal.pgen.1005241)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Hashimoto S, Morimoto N, Yamanaka M, Matsumoto H, Yamochi T, Goto H, Inoue M, Nakaoka Y, Shibahara H & & Morimoto Y 2017 Quantitative and qualitative changes of mitochondria in human preimplantation embryos. Journal of Assisted Reproduction and Genetics 34 573–580. (https://doi.org/10.1007/s10815-017-0886-6)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Held-Hoelker E, Klein SL, Rings F, Salilew-Wondim D, Saeed-Zidane M, Neuhoff C, Tesfaye D, Schellander K & & Hoelker M 2017 Cryosurvival of in vitro produced bovine embryos supplemented with l-carnitine and concurrent reduction of fatty acids. Theriogenology 96 145–152. (https://doi.org/10.1016/j.theriogenology.2017.03.014)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Holm P, Booth PJ, Schmidt MH, Greve T & & Callesen H 1999 High bovine blastocyst development in a static in vitro production system using SOFaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins. Theriogenology 52 683–700. (https://doi.org/10.1016/S0093-691X(9900162-4)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Itami N, Shirasuna K, Kuwayama T & & Iwata H 2015 Resveratrol improves the quality of pig oocytes derived from early antral follicles through sirtuin 1 activation. Theriogenology 83 1360–1367. (https://doi.org/10.1016/j.theriogenology.2015.01.029)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Ji J, Chen L, Zhuang Y, Han Y, Tang W & & Xia F 2020 Fibronectin 1 inhibits the apoptosis of human trophoblasts by activating the PI3K/Akt signaling pathway. International Journal of Molecular Medicine 46 1908–1922. (https://doi.org/10.3892/ijmm.2020.4735)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Jones R, Peña J, Mystal E, Marsit C, Lee M-J, Stone J & & Lambertini L 2020 Mitochondrial and glycolysis-regulatory gene expression profiles are associated with intrauterine growth restriction. Journal of Maternal-Fetal and Neonatal Medicine 33 1336–1345. (https://doi.org/10.1080/14767058.2018.1518419)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Konstantinidis M, Alfarawati S, Hurd D, Paolucci M, Shovelton J, Fragouli E & & Wells D 2014 Simultaneous assessment of aneuploidy, polymorphisms, and mitochondrial DNA content in human polar bodies and embryos with the use of a novel microarray platform. Fertility and Sterility 102 1385–1392. (https://doi.org/10.1016/j.fertnstert.2014.07.1233)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Krisher RL & & Prather RS 2012 A role for the Warburg effect in preimplantation embryo development: metabolic modification to support rapid cell proliferation. Molecular Reproduction and Development 79 311–320. (https://doi.org/10.1002/mrd.22037)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Kurzella J, Miskel D, Rings F, Tholen E, Tesfaye D, Schellander K, Salilew-Wondim D, Held-Hoelker E, Große-Brinkhaus C & & Hoelker M 2023 The mitochondrial respiration signature of the bovine blastocyst reflects both environmental conditions of development as well as embryo quality. Scientific Reports 13 19408. (https://doi.org/10.1038/s41598-023-45691-2)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Le Zhao YY, Yang H, Luo N, Li X, Zheng J, Yang S & & Zhao Y 2023 Screening genes related to embryo implantation in Dazu black goats (Capra hircus) by morphological and transcriptome analyses. Journal of Animal Science 101. (https://doi.org/10.1016/j.ebiom.2019.10.018)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Lee SH, Liu X, Jimenez-Morales D & & Rinaudo PF 2022 Murine blastocysts generated by in vitro fertilization show increased Warburg metabolism and altered lactate production. eLife 11. (https://doi.org/10.7554/eLife.79153)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Leese HJ, Guerif F, Allgar V, Brison DR, Lundin K & & Sturmey RG 2016 Biological optimization, the Goldilocks principle, and how much is lagom in the preimplantation embryo. Molecular Reproduction and Development 83 748–754. (https://doi.org/10.1002/mrd.22684)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Leme LO, Machado GM, Fidelis AAG, Guimarães ALS, Sprícigo JFW, Carvalho JO, Pivato I, Franco MM & & Dode MAN 2021 Transcriptome of D14 in vivo x in vitro bovine embryos: is there any difference? In Vitro Cellular & Developmental Biology. Animal 57 598–609. (https://doi.org/10.1007/s11626-021-00599-x)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Li J, Liu L, Weng J, Yin T-L, Yang J & & Feng HL 2021 Biological roles of L-carnitine in oocyte and early embryo development. Molecular Reproduction and Development 88 673–685. (https://doi.org/10.1002/mrd.23542)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Lim H & & Dey SK 2000 PPAR delta functions as a prostacyclin receptor in blastocyst implantation. Trends in Endocrinology and Metabolism: TEM 11 137–142. (https://doi.org/10.1016/s1043-2760(0000243-5)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Longo N, Frigeni M & & Pasquali M 2016 Carnitine transport and fatty acid oxidation. Biochimica et Biophysica Acta 1863 2422–2435. (https://doi.org/10.1016/j.bbamcr.2016.01.023)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • May-Panloup P, Boguenet M, Hachem HE, Bouet P-E & & Reynier P 2021 Embryo and its mitochondria. Antioxidants 10. (https://doi.org/10.3390/antiox10020139)

  • Merviel P, Challier JC, Carbillon L, Foidart JM & & Uzan S 2001 The role of integrins in human embryo implantation. Fetal Diagnosis and Therapy 16 364–371. (https://doi.org/10.1159/000053942)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Moore R, Tao W, Smith ER & & Xu X-X 2014 The primitive endoderm segregates from the epiblast in β1 integrin-deficient early mouse embryos. Molecular and Cellular Biology 34 560–572. (https://doi.org/10.1128/MCB.00937-13)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Noguchi T, Aizawa T, Munakata Y & & Iwata H 2020 Comparison of gene expression and mitochondria number between bovine blastocysts obtained in vitro and in vivo. Journal of Reproduction and Development 66 35–39. (https://doi.org/10.1262/jrd.2019-100)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Papis K, Poleszczuk O, Wenta-Muchalska E & & Modlinski JA 2007 Melatonin effect on bovine embryo development in vitro in relation to oxygen concentration. Journal of Pineal Research 43 321–326. (https://doi.org/10.1111/j.1600-079X.2007.00479.x)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Parrish JJ 2014 Bovine in vitro fertilization: in vitro oocyte maturation and sperm capacitation with heparin. Theriogenology 81 67–73. (https://doi.org/10.1016/j.theriogenology.2013.08.005)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Pasquariello R, Zhang M, Herrick JR, Ermisch A, Becker J, Schoolcraft WB, Barfield JP, Yuan Y & & Krisher RL 2023 Lipid enriched reduced nutrient culture medium improves bovine blastocyst formation. Reproduction and Fertility 4. (https://doi.org/10.1530/RAF-23-0057)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Pavani KC, Rocha A, Oliveira E, da Silva FM & & Sousa M 2020 Novel ultrastructural findings in bovine oocytes matured in vitro. Theriogenology 143 88–97. (https://doi.org/10.1016/j.theriogenology.2019.12.003)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Placidi M, Di Emidio G, Virmani A, D'Alfonso A, Artini PG, D'Alessandro AM & & Tatone C 2022 Carnitines as mitochondrial modulators of oocyte and embryo bioenergetics. Antioxidants 11. (https://doi.org/10.3390/antiox11040745)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Quijano C, Trujillo M, Castro L & & Trostchansky A 2016 Interplay between oxidant species and energy metabolism. Redox Biology 8 28–42. (https://doi.org/10.1016/j.redox.2015.11.010)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Ribeiro ES, Greco LF, Bisinotto RS, Lima FS, Thatcher WW & & Santos JE 2016 Biology of preimplantation conceptus at the onset of elongation in dairy cows. Biology of Reproduction 94 97. (https://doi.org/10.1095/biolreprod.115.134908)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Robinson MD, McCarthy DJ & & Smyth GK 2010 edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26 139–140. (https://doi.org/10.1093/bioinformatics/btp616)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Sánchez JM, Passaro C, Forde N, Browne JA, Behura SK, Fernández-Fuertes B, Mathew DJ, Kelly AK, Butler ST & Spencer TE et al.2019 Do differences in the endometrial transcriptome between uterine horns ipsilateral and contralateral to the corpus luteum influence conceptus growth to day 14 in cattle? Biology of Reproduction 100 86–100. (https://doi.org/10.1093/biolre/ioy185)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Santos JEP, Thatcher WW, Chebel RC, Cerri RLA & & Galvão KN 2004 The effect of embryonic death rates in cattle on the efficacy of estrus synchronization programs. Animal Reproduction Science 82–83 513–535. (https://doi.org/10.1016/j.anireprosci.2004.04.015)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Seidel GE 2006 Modifying oocytes and embryos to improve their cryopreservation. Theriogenology 65 228–235. (https://doi.org/10.1016/j.theriogenology.2005.09.025)

  • Sturmey RG & & Leese HJ 2003 Energy metabolism in pig oocytes and early embryos. Reproduction 126 197–204. (https://doi.org/10.1530/rep.0.1260197)

  • Takahashi T, Inaba Y, Somfai T, Kaneda M, Geshi M, Nagai T & & Manabe N 2013 Supplementation of culture medium with L-carnitine improves development and cryotolerance of bovine embryos produced in vitro. Reproduction, Fertility, and Development 25 589–599. (https://doi.org/10.1071/RD11262)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Tríbulo P, Rabaglino MB, Bo MB, Carvalheira LdR, Bishop JV, Hansen TR & & Hansen PJ 2019 Dickkopf-related protein 1 is a progestomedin acting on the bovine embryo during the morula-to-blastocyst transition to program trophoblast elongation. Scientific Reports 9 11816. (https://doi.org/10.1038/s41598-019-48374-z)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Trimarchi JR, Liu L, Porterfield DM, Smith PJ & & Keefe DL 2000 Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos. Biology of Reproduction 62 1866–1874. (https://doi.org/10.1095/biolreprod62.6.1866)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Truong TT, Soh YM & & Gardner DK 2016 Antioxidants improve mouse preimplantation embryo development and viability. Human Reproduction 31 1445–1454. (https://doi.org/10.1093/humrep/dew098)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Wiltbank MC, Souza AH, Carvalho PD, Cunha AP, Giordano JO, Fricke PM, Baez GM & & Diskin MG 2014 Physiological and practical effects of progesterone on reproduction in dairy cattle. Animal 8(Supplement 1) 70–81. (https://doi.org/10.1017/S1751731114000585)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Yan S, Wang Y, Chen M, Li G & & Fan J 2015 Deregulated SLC2A1 promotes tumor cell proliferation and metastasis in gastric cancer. International Journal of Molecular Sciences 16 16144–16157. (https://doi.org/10.3390/ijms160716144)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Yoon S-B, Choi S-A, Sim B-W, Kim J-S, Mun S-E, Jeong P-S, Yang H-J, Lee Y, Park Y-H & Song B-S et al.2014 Developmental competence of bovine early embryos depends on the coupled response between oxidative and endoplasmic reticulum stress. Biology of Reproduction 90 104. (https://doi.org/10.1095/biolreprod.113.113480)

    • PubMed
    • Search Google Scholar
    • Export Citation

  • Young LE, Sinclair KD & & Wilmut I 1998 Large offspring syndrome in cattle and sheep. Reviews of Reproduction 3 155–163. (https://doi.org/10.1530/ror.0.0030155)

  • Zolini AM, Carrascal-Triana E, Ruiz de King A, Hansen PJ, Alves Torres CA & & Block J 2019 Effect of addition of L-carnitine to media for oocyte maturation and embryo culture on development and cryotolerance of bovine embryos produced in vitro. Theriogenology 133 135–143. (https://doi.org/10.1016/j.theriogenology.2019.05.005)

    • PubMed
    • Search Google Scholar
    • Export Citation