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Dawit Tesfaye Institute of Animal Sciences, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany
Center of Integrated Dairy Research, University of Bonn, Bonn, Germany

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Samuel Gebremedhn Institute of Animal Sciences, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany
Center of Integrated Dairy Research, University of Bonn, Bonn, Germany

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Dessie Salilew-Wondim Institute of Animal Sciences, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany
Center of Integrated Dairy Research, University of Bonn, Bonn, Germany

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Tsige Hailay Institute of Animal Sciences, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany
Center of Integrated Dairy Research, University of Bonn, Bonn, Germany

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Michael Hoelker Institute of Animal Sciences, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany
Center of Integrated Dairy Research, University of Bonn, Bonn, Germany

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Christine Grosse-Brinkhaus Institute of Animal Sciences, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany

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Karl Schellander Institute of Animal Sciences, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany
Center of Integrated Dairy Research, University of Bonn, Bonn, Germany

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The genetic regulation of female fertility (follicular development, oocyte maturation and early preimplantation embryo development) involves the spatio-temporal regulation of those genes that play key roles in various stages of the female reproductive axis. MicroRNAs (miRNAs), a class of small non-coding RNAs, are known to regulate the expression of a large proportion of such genes. In recent decades, multiple studies have aimed to determine the roles of these non-coding RNAs in mammalian follicular development, oocyte growth and embryo development. These studies have applied a variety of approaches, including conditional knockout of miRNA biogenesis genes, high-throughput sequencing technologies for pattern recognition in miRNA expression and loss- and gain-of-function of miRNAs in various animal models. In addition to the cellular miRNAs, a large variety of RNAs are found in circulation, being coupled with extracellular vesicles, proteins and lipids. Because of their potential as diagnostic markers for abnormal physiologies, there is increasing interest in the identification of extracellular miRNAs in various biological fluids and spent in vitro culture media. This review focuses on studies addressing the expression and potential role of cellular and extracellular miRNAs in mammalian follicular cell physiology and subsequent ovarian functionality and oocyte maturation.

Article Type:
Review Article
DOI:
https://doi.org/10.1530/REP-17-0428
Volume/Issue:
Volume 155: Issue 3
Free access
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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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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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.

Article Type:
Research Article
DOI:
https://doi.org/10.1530/REP-24-0004
Volume/Issue:
Volume 168: Issue 3
Restricted access
Salilew-Wondim Dessie Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Franca Rings Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Michael Hölker Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Markus Gilles Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Danyel Jennen Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Ernst Tholen Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Vitezslav Havlicek Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Urban Besenfelder Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Vladimir L Sukhorukov Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Ulrich Zimmermann Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Joerg M Endter Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Marc-André Sirard Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Karl Schellander Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Dawit Tesfaye Animal Breeding and Husbandry Group, Institute of Animal Science, University of Bonn, Endenicher Allee 15, Bonn, Germany, University of Veterinary Medicine, Veterinär platz 1, A-1210 Vienna, Austria, Biotechnology center, University of Wuerzburg, D-97074, Wuerzburg, Germany and Départment des Science Animales, Centre de Recherche en Biologie de la Reproduction, Université Laval, G1K7p4, Quebec, Canada

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Selecting developmentally competent oocytes and zygotes based on their morphology is more often influenced by personal judgments and lacks universal standards. Therefore, this experiment was conducted to investigate the rate of development and mRNA level of dielectrophoretically separated oocytes and zygotes to validate dielectrophoresis (DEP) as non-invasive option for selection of oocytes and zygotes. In the first experiment, metaphase II oocytes with (PB+) and without (PB−) first polar body and zygotes were subjected to DEP at 4 MHz and 450 μm electrode distance and classified into fast, very fast, slow, and very slow depending on the time elapsed to reach one of the electrodes in the electric field. Parthenogenetic activation was employed to monitor the embryonic development of dielectrophoretically classified oocytes. The result revealed that at 6 and 7 days of post-activation, the blastocyst rate of very slow dielectrophoretic PB+ and PB− oocytes was significantly (P < 0.05) lower than other groups. Similarly, in zygotes, the blastocyst rate at 7 days post-insemination was higher (P < 0.05) in the very fast dielectrophoretic categories when compared with the slow and very slow categories. In the second experiment, mRNA level was analyzed in the very fast and very slow dielectrophoretic PB+ oocytes and zygotes respectively using the bovine cDNA microarray. The result showed that 36 and 42 transcripts were differentially regulated between the very fast and very slow dielectrophoretic categories PB+ oocytes and zygotes respectively. In conclusion, dielectrophoretically separated oocytes and zygotes showed difference in the rate of blastocyst development accompanied by difference in transcriptional abundances.

Article Type:
Research Article
DOI:
https://doi.org/10.1530/REP-06-0277
Volume/Issue:
Volume 133: Issue 5
Free access