Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Differential gene expression in the endometrium reveals cytoskeletal and immunological genes in lactating dairy cows genetically divergent for fertility traits

Bruce Moran A C , Stephen T. Butler B F , Stephen G. Moore B C , David E. MacHugh C D and Christopher J. Creevey A E
+ Author Affiliations
- Author Affiliations

A Teagasc, Animal & Grassland Research and Innovation Centre, Grange, Dunsany, Co. Meath, Ireland.

B Teagasc, Animal & Grassland Research and Innovation Centre, Moorepark, Fermoy, Co. Cork, Ireland.

C Animal Genomics Laboratory, UCD School of Agriculture and Food Science, University College Dublin, Belfield, Dublin 4, Ireland.

D UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.

E Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, SY23 3FG, UK.

F Corresponding author. Email: stephen.butler@teagasc.ie

Reproduction, Fertility and Development 29(2) 274-282 https://doi.org/10.1071/RD15128
Submitted: 1 April 2015  Accepted: 27 June 2015   Published: 25 August 2015

Abstract

Profitable milk production in dairy cows requires good reproductive performance. Calving interval is a trait used to measure reproductive efficiency. Herein we used a novel lactating Holstein cow model of fertility that displayed genetic and phenotypic divergence in calving interval, a trait used to define reproductive performance using a national breeding index in Ireland. Cows had similar genetic merit for milk production traits, but either very good genetic merit for fertility (Fert+; n = 7) or very poor genetic merit for fertility (Fert–; n = 6). We tested the hypothesis that Fert+ cows would have a corresponding detectable difference in endometrial gene expression compared with the Fert– cows. To do this, we sequenced the transcriptome of endometrial biopsies collected on Day 7 of the oestrous cycle (non-pregnant). This is an important stage for uterine remodelling and initiation of histotroph secretion. Significant differential expression (false discovery rate-adjusted P < 0.1) of 403 genes between Fert+ and Fert– cows was found. A novel network-based functional analysis highlighted 123 genes from three physiologically relevant networks of the endometrium: (1) actin and cytoskeletal components; (2) immune function; and (3) ion transportation. In particular, our results indicate an overall downregulation of inflammation-related genes and an upregulation of multiple ion transporters and gated-voltage channels and cytoskeletal genes in Fert+ cows. These three topics, which are discussed in terms of the uterus and in the context of fertility, provide molecular evidence for an association between gene expression in the uterine environment and genetic merit for fertility in dairy cows.

Additional keywords: fertility traits, genetic selection, reproduction, transcriptome, uterine environment, uterus.


References

Aguilar, H. N., and Mitchell, B. F. (2010). Physiological pathways and molecular mechanisms regulating uterine contractility. Hum. Reprod. Update 16, 725–744.
Physiological pathways and molecular mechanisms regulating uterine contractility.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhtlShu7%2FP&md5=1b058f2d5d7009b6081641fb579673ebCAS | 20551073PubMed |

Ahn, H. W., Farmer, J. L., Bazer, F. W., and Spencer, T. E. (2009). Progesterone and interferon tau-regulated genes in the ovine uterine endometrium: identification of periostin as a potential mediator of conceptus elongation. Reproduction 138, 813–825.
Progesterone and interferon tau-regulated genes in the ovine uterine endometrium: identification of periostin as a potential mediator of conceptus elongation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhsValt7rE&md5=5e371e93df5e4b8f69a72fd7aa7986c2CAS | 19635739PubMed |

Anders, S. (2010). Counting reads in features: HTSeq. (EMBL, Huber Group: Heidelberg, Germany.) Available at: http://www-huber.embl.de/users/anders/HTSeq/doc/count.html [verified 29 October 2013].

Andrews, S. (2013). FastQC: a quality control tool for high throughput sequence data. (Babraham Institute: Cambridge, UK). Available at: http://www.bioinformatics.babraham.ac.uk/projects/fastqc/ [verified 29 October 2013].

Bastian, M., Heymann, S., and Jacomy, M. (2009). Gephi: an open source software for exploring and manipulating networks. Int. AAAI Conf. Weblogs Soc. Media. Available at https://gephi.org/publications/gephi-bastian-feb09.pdf [Verified 24 July 2015]

Bauersachs, S. (2005). Gene expression profiling of bovine endometrium during the oestrous cycle: detection of molecular pathways involved in functional changes. J. Mol. Endocrinol. 34, 889–908.
Gene expression profiling of bovine endometrium during the oestrous cycle: detection of molecular pathways involved in functional changes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXlslCisL8%3D&md5=1968a65b71fdbb8146a0ba5209e94b12CAS | 15956356PubMed |

Bauersachs, S. (2006). Embryo-induced transcriptome changes in bovine endometrium reveal species-specific and common molecular markers of uterine receptivity. Reproduction 132, 319–331.
Embryo-induced transcriptome changes in bovine endometrium reveal species-specific and common molecular markers of uterine receptivity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xpt1Wjs7s%3D&md5=564470651c7589b3eaafdcceb1318523CAS | 16885540PubMed |

Bauersachs, S., and Wolf, E. (2012). Transcriptome analyses of bovine, porcine and equine endometrium during the pre-implantation phase. Anim. Reprod. Sci. 134, 84–94.
Transcriptome analyses of bovine, porcine and equine endometrium during the pre-implantation phase.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38Xht1ekurrP&md5=7c279178f2a3af8e05b2c5f2c3a873c7CAS | 22917876PubMed |

Benjamini, Y., and Hochberg, Y. (1995). Contolling the false discovery rate: a practical and powerful approach to multiple testing. Available at: http://www.jstor.org.eproxy.ucd.ie/stable/2346101 [verified 30 May 2013].

Berry, D. P., Buckley, F., Dillon, P., Evans, R. D., Rath, M., and Veerkamp, R. F. (2003). Genetic parameters for body condition score, body weight, milk yield, and fertility estimated using random regression models. J. Dairy Sci. 86, 3704–3717.
Genetic parameters for body condition score, body weight, milk yield, and fertility estimated using random regression models.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXptFaisr8%3D&md5=98809066388a27332402be431310d4d7CAS | 14672201PubMed |

Butler, S. T. (2014). Genetic control of reproduction in dairy cows. Reprod. Fertil. Dev. 26, 1–11.
Genetic control of reproduction in dairy cows.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhvV2lsrrK&md5=963563e13a7730db43d32b9671b457dcCAS |

Callegari, E. A., Ferguson-Gottschall, S., and Gibori, G. (2005). PGF2alpha induced differential expression of genes involved in turnover of extracellular matrix in rat decidual cells. Reprod. Biol. Endocrinol. 3, 3.
PGF2alpha induced differential expression of genes involved in turnover of extracellular matrix in rat decidual cells.Crossref | GoogleScholarGoogle Scholar | 15644143PubMed |

Carp, H. J. A., Selmi, C., and Shoenfeld, Y. (2012). The autoimmune bases of infertility and pregnancy loss. J. Autoimmun. 38, J266–J274.
The autoimmune bases of infertility and pregnancy loss.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XktlGkuro%3D&md5=3c9765741445e7643523bee2465d4d8cCAS |

Chapwanya, A., Meade, K. G., Narciandi, F., Stanley, P., Mee, J. F., Doherty, M. L., Callanan, J. J., and O’Farrelly, C. (2010). Endometrial biopsy: a valuable clinical and research tool in bovine reproduction. Theriogenology 73, 988–994.
Endometrial biopsy: a valuable clinical and research tool in bovine reproduction.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3c3gsVSlsg%3D%3D&md5=62acce532a396df526b4f2c314b2a749CAS | 20083303PubMed |

Christensen, S., Verhage, H. G., Nowak, G., de Lanerolle, P., Fleming, S., Bell, S. C., Fazleabas, A. T., and Hild-Petito, S. (1995). Smooth muscle myosin II and alpha smooth muscle actin expression in the baboon (Papio anubis) uterus is associated with glandular secretory activity and stromal cell transformation. Biol. Reprod. 53, 598–608.
Smooth muscle myosin II and alpha smooth muscle actin expression in the baboon (Papio anubis) uterus is associated with glandular secretory activity and stromal cell transformation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXns1ylsLw%3D&md5=ceb5093e0aa2721de89dee4c67a6d4acCAS | 7578684PubMed |

Cortesio, C. L., and Jiang, W. (2006). Mannan-binding lectin-associated serine protease 3 cleaves synthetic peptides and insulin-like growth factor-binding protein 5. Arch. Biochem. Biophys. 449, 164–170.
Mannan-binding lectin-associated serine protease 3 cleaves synthetic peptides and insulin-like growth factor-binding protein 5.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XktVyhtLw%3D&md5=d12487f84d55e5daf1e0706cc8bbd038CAS | 16554018PubMed |

Coutts, A. S., MacKenzie, E., Griffith, E., and Black, D. M. (2003). TES is a novel focal adhesion protein with a role in cell spreading. J. Cell Sci. 116, 897–906.
TES is a novel focal adhesion protein with a role in cell spreading.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXitlOju7w%3D&md5=e92003374e7d0bead56b5d2fb0be91eeCAS | 12571287PubMed |

Cummins, S. B., Lonergan, P., Evans, A. C. O., Berry, D. P., Evans, R. D., and Butler, S. T. (2012a). Genetic merit for fertility traits in Holstein cows: I. Production characteristics and reproductive efficiency in a pasture-based system. J. Dairy Sci. 95, 1310–1322.
Genetic merit for fertility traits in Holstein cows: I. Production characteristics and reproductive efficiency in a pasture-based system.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XjtFWgurY%3D&md5=b45ba92b7064e999b6d7ef61ce86c388CAS | 22365213PubMed |

Cummins, S. B., Lonergan, P., Evans, A. C. O., and Butler, S. T. (2012b). Genetic merit for fertility traits in Holstein cows: II. Ovarian follicular and corpus luteum dynamics, reproductive hormones, and estrus behavior. J. Dairy Sci. 95, 3698–3710.
Genetic merit for fertility traits in Holstein cows: II. Ovarian follicular and corpus luteum dynamics, reproductive hormones, and estrus behavior.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XoslGnsrc%3D&md5=8b445ae2d87be2bb2cffe57efa450ff8CAS | 22720927PubMed |

Cummins, S. B., Waters, S. M., Evans, A. C. O., Lonergan, P., and Butler, S. T. (2012c). Genetic merit for fertility traits in Holstein cows: III. Hepatic expression of somatotropic axis genes during pregnancy and lactation. J. Dairy Sci. 95, 3711–3721.
Genetic merit for fertility traits in Holstein cows: III. Hepatic expression of somatotropic axis genes during pregnancy and lactation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XoslGrt74%3D&md5=610cdd5cabea991b2a0c442be0c5ddbaCAS | 22720928PubMed |

Dobin, A., Davis, C. A., Schlesinger, F., Drenkow, J., Zaleski, C., Jha, S., Batut, P., Chaisson, M., and Gingeras, T. R. (2013). STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29, 15–21.
STAR: ultrafast universal RNA-seq aligner.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhvV2gsbnF&md5=e9f48f79de5fc7dd4b8315ea869bcd72CAS | 23104886PubMed |

Durinck, S., Spellman, P. T., Birney, E., and Huber, W. (2009). Mapping identifiers for the integration of genomic datasets with the R/Bioconductor package biomaRt. Nat. Protoc. 4, 1184–1191.
Mapping identifiers for the integration of genomic datasets with the R/Bioconductor package biomaRt.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXpt12ksrk%3D&md5=7a162956f3d5ccc522fc51f8541f090eCAS | 19617889PubMed |

ENSEMBL (2013). ‘Bos taurus UMD 3.1.’ Available at: http://www.ensembl.org/info/data/ftp/index.html [verified 29 October 2013].

Evans, A. C. O., and Walsh, S. W. (2012). The physiology of multifactorial problems limiting the establishment of pregnancy in dairy cattle. Reprod. Fertil. Dev. 24, 233–237.
The physiology of multifactorial problems limiting the establishment of pregnancy in dairy cattle.Crossref | GoogleScholarGoogle Scholar |

Evans, R. D., Wallace, M., Shalloo, L., Garrick, D. J., and Dillon, P. (2006). Financial implications of recent declines in reproduction and survival of Holstein–Friesian cows in spring-calving Irish dairy herds. Agric. Syst. 89, 165–183.
Financial implications of recent declines in reproduction and survival of Holstein–Friesian cows in spring-calving Irish dairy herds.Crossref | GoogleScholarGoogle Scholar |

Forde, N., and Lonergan, P. (2012). Transcriptomic analysis of the bovine endometrium: what is required to establish uterine receptivity to implantation in cattle? J. Reprod. Dev. 58, 189–195.
Transcriptomic analysis of the bovine endometrium: what is required to establish uterine receptivity to implantation in cattle?Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XptVKju7Y%3D&md5=cc0cb8b2b87283073b66dabe33bc6c4cCAS | 22738902PubMed |

Forde, N., Carter, F., Fair, T., Crowe, M. A., Evans, A. C. O., Spencer, T. E., Bazer, F. W., McBride, R., Boland, M. P., O’Gaora, P., Lonergan, P., and Roche, J. F. (2009). Progesterone-regulated changes in endometrial gene expression contribute to advanced conceptus development in cattle. Biol. Reprod. 81, 784–794.
Progesterone-regulated changes in endometrial gene expression contribute to advanced conceptus development in cattle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhtFyhsLbM&md5=fdff0efe89c1877c45b80279b217564cCAS | 19553605PubMed |

Forde, N., Spencer, T. E., Bazer, F. W., Song, G., Roche, J. F., and Lonergan, P. (2010). Effect of pregnancy and progesterone concentration on expression of genes encoding for transporters or secreted proteins in the bovine endometrium. Physiol. Genomics 41, 53–62.
Effect of pregnancy and progesterone concentration on expression of genes encoding for transporters or secreted proteins in the bovine endometrium.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhtlakt7jN&md5=d4dd53564ea65fd464424c724f18e70bCAS | 19996158PubMed |

Forde, N., Carter, F., Spencer, T. E., Bazer, F. W., Sandra, O., Mansouri-Attia, N., Okumu, L. A., McGettigan, P. A., Mehta, J. P., McBride, R., O’Gaora, P., Roche, J. F., and Lonergan, P. (2011). Conceptus-induced changes in the endometrial transcriptome: how soon does the cow know she is pregnant? Biol. Reprod. 85, 144–156.
Conceptus-induced changes in the endometrial transcriptome: how soon does the cow know she is pregnant?Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXotFOlsL4%3D&md5=f72f2579f7206315fe155fea0de513fdCAS | 21349821PubMed |

Forde, N., Duffy, G. B., McGettigan, P. A., Browne, J. A., Mehta, J. P., Kelly, A. K., Mansouri-Attia, N., Sandra, O., Loftus, B. J., Crowe, M. A., Fair, T., Roche, J. F., Lonergan, P., and Evans, A. C. (2012a). Evidence for an early endometrial response to pregnancy in cattle: both dependent upon and independent of interferon tau. Physiol. Genomics 44, 799–810.
Evidence for an early endometrial response to pregnancy in cattle: both dependent upon and independent of interferon tau.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhslamuw%3D%3D&md5=1ccaf9ba21f6e914cf4b94c5342c23d1CAS | 22759920PubMed |

Forde, N., Mehta, J. P., Minten, M., Crowe, M. A., Roche, J. F., Spencer, T. E., and Lonergan, P. (2012b). Effects of low progesterone on the endometrial transcriptome in cattle. Biol. Reprod. 87, 124.
Effects of low progesterone on the endometrial transcriptome in cattle.Crossref | GoogleScholarGoogle Scholar | 23018184PubMed |

Forde, N., Mehta, J. P., McGettigan, P. A., Mamo, S., Bazer, F. W., Spencer, T. E., and Lonergan, P. (2013). Alterations in expression of endometrial genes coding for proteins secreted into the uterine lumen during conceptus elongation in cattle. BMC Genomics 14, 321.
Alterations in expression of endometrial genes coding for proteins secreted into the uterine lumen during conceptus elongation in cattle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXptlyiu7w%3D&md5=db94f9d02839be0a1fd1a89669953ebbCAS | 23663413PubMed |

Groebner, A. E., Rubio-Aliaga, I., Schulke, K., Reichenbach, H. D., Daniel, H., Wolf, E., Meyer, H. H., and Ulbrich, S. E. (2011). Increase of essential amino acids in the bovine uterine lumen during preimplantation development. Reproduction 141, 685–695.
Increase of essential amino acids in the bovine uterine lumen during preimplantation development.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXmvFCis74%3D&md5=33a05192480161612972a0e84ad2cbd5CAS | 21383026PubMed |

Gunning, P., O’neill, G., and Hardeman, E. (2008). Tropomyosin-based regulation of the actin cytoskeleton in time and space. Physiol. Rev. 88, 1–35.
Tropomyosin-based regulation of the actin cytoskeleton in time and space.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXitVKgsbg%3D&md5=3a5b58d5c9e053455999e3570a3b8df3CAS | 18195081PubMed |

Haller-Kikkatalo, K., Salumets, A., and Uibo, R. (2012). Review on autoimmune reactions in female infertility: antibodies to follicle stimulating hormone. Clin. Dev. Immunol. 2012, Article ID 762541.
Review on autoimmune reactions in female infertility: antibodies to follicle stimulating hormone.Crossref | GoogleScholarGoogle Scholar |

Herlihy, M. M., Berry, D. P., Crowe, M. A., Diskin, M. G., and Butler, S. T. (2011). Evaluation of protocols to synchronize estrus and ovulation in seasonal calving pasture-based dairy production systems. J. Dairy Sci. 94, 4488–4501.
Evaluation of protocols to synchronize estrus and ovulation in seasonal calving pasture-based dairy production systems.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhtV2gsLbP&md5=5cc0cd8cba7c972093be7df8e22a612aCAS | 21854921PubMed |

Hoekstra, J., van der Lugt, A., van der Werf, J. H., and Ouweltjes, W. (1994). Genetic and phenotypic parameters for milk production and fertility traits in upgraded dairy cattle. Livest. Prod. Sci. 40, 225–232.
Genetic and phenotypic parameters for milk production and fertility traits in upgraded dairy cattle.Crossref | GoogleScholarGoogle Scholar |

Huang, D. W., Sherman, B. T., and Lempicki, R. A. (2009a). Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res. 37, 1–13.
Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists.Crossref | GoogleScholarGoogle Scholar |

Huang, D. W., Sherman, B. T., and Lempicki, R. A. (2009b). Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat. Protoc. 4, 44–57.
Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsFCkurnI&md5=e95521288c9064241695903dc4556814CAS |

Johnson, G. A., Burghardt, R. C., Joyce, M. M., Spencer, T. E., Bazer, F. W., Pfarrer, C., and Gray, C. A. (2003a). Osteopontin expression in uterine stroma indicates a decidualization-like differentiation during ovine pregnancy. Biol. Reprod. 68, 1951–1958.
Osteopontin expression in uterine stroma indicates a decidualization-like differentiation during ovine pregnancy.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXks1Ggsrk%3D&md5=c6865ea2c31c768ad526a03e8a34f77aCAS | 12606396PubMed |

Johnson, G. A., Burghardt, R. C., Bazer, F. W., and Spencer, T. E. (2003b). Osteopontin: roles in implantation and placentation. Biol. Reprod. 69, 1458–1471.
Osteopontin: roles in implantation and placentation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXosV2kur8%3D&md5=225143da285298e6cce7074f81742f71CAS | 12890718PubMed |

Khurana, S., and George, S. P. (2008). Regulation of cell structure and function by actin-binding proteins: villin’s perspective. FEBS Lett. 582, 2128–2139.
Regulation of cell structure and function by actin-binding proteins: villin’s perspective.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXms1Wgtbs%3D&md5=db2ab9a028e30444a220239b60c50ec1CAS | 18307996PubMed |

Korgun, E. T., Cayli, S., Asar, M., and Demir, R. (2007). Distribution of laminin, vimentin and desmin in the rat uterus during initial stages of implantation. J. Mol. Histol. 38, 253–260.
Distribution of laminin, vimentin and desmin in the rat uterus during initial stages of implantation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXotFajsb4%3D&md5=606f6c8360d54affe321418ab2065c1cCAS | 17653607PubMed |

LeBleu, V. S., Macdonald, B., and Kalluri, R. (2007). Structure and function of basement membranes. Exp. Biol. Med. (Maywood) 232, 1121–1129.
Structure and function of basement membranes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhtFKnurjF&md5=1f9e4d242cd2fca774f6f62f7bb57f03CAS | 17895520PubMed |

Lee, B. P., Rushlow, W. J., Chakraborty, C., and Lala, P. K. (2001). Differential gene expression in premalignant human trophoblast: role of IGFBP-5. Int. J. Cancer 94, 674–684.
Differential gene expression in premalignant human trophoblast: role of IGFBP-5.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXotFalurw%3D&md5=89371371b370b3ff2149d52dd965e181CAS | 11745462PubMed |

Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., Homer, N., Marth, G., Abecasis, G., Durbin, R., 1000 Genome Project Data Processing Subgroup (2009). The Sequence Alignment/Map format and SAMtools. Bioinformatics 25, 2078–2079.
The Sequence Alignment/Map format and SAMtools.Crossref | GoogleScholarGoogle Scholar | 19505943PubMed |

McCabe, M., Waters, S., Morris, D., Kenny, D., Lynn, D., and Creevey, C. (2012). RNA-seq analysis of differential gene expression in liver from lactating dairy cows divergent in negative energy balance. BMC Genomics 13, 193.
RNA-seq analysis of differential gene expression in liver from lactating dairy cows divergent in negative energy balance.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhslGqu7rL&md5=71e0f22f48ea85c7ad06891d00e97027CAS | 22607119PubMed |

Mitko, K., Ulbrich, S. E., Wenigerkind, H., Sinowatz, F., Blum, H., Wolf, E., and Bauersachs, S. (2008). Dynamic changes in messenger RNA profiles of bovine endometrium during the oestrous cycle. Reproduction 135, 225–240.
Dynamic changes in messenger RNA profiles of bovine endometrium during the oestrous cycle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXit1yrtLo%3D&md5=3609a6ec7819f691e79ef77f35f79fb4CAS | 18239051PubMed |

Moore, S. G., Fair, T., Lonergan, P., and Butler, S. T. (2014a). Genetic merit for fertility traits in Holstein cows: IV. Transition period, uterine health, and resumption of cyclicity. J. Dairy Sci. 97, 2740–2752.
Genetic merit for fertility traits in Holstein cows: IV. Transition period, uterine health, and resumption of cyclicity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXktlWkurk%3D&md5=0844130f64bd701c8ea98b517dc42429CAS | 24630663PubMed |

Moore, S. G., Scully, S., Browne, J. A., and Butler, S. T. (2014b). Genetic merit for fertility traits in Holstein cows: V. Factors affecting circulating progesterone concentrations. J. Dairy Sci. 97, 5543–5557.
Genetic merit for fertility traits in Holstein cows: V. Factors affecting circulating progesterone concentrations.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXhtVCjur3J&md5=2cc2781ac26403aad40b97f1c0892174CAS | 24952779PubMed |

Mortazavi, A., Williams, B. A., McCue, K., Schaeffer, L., and Wold, B. (2008). Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat. Methods 5, 621–628.
Mapping and quantifying mammalian transcriptomes by RNA-Seq.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXnslyqs7k%3D&md5=0fd254b12f8780b5a830bdfd2446584cCAS | 18516045PubMed |

Nakamura, F., Stossel, T. P., and Hartwig, J. H. (2011). The filamins: organizers of cell structure and function. Cell Adh. Migr. 5, 160–169.
The filamins: organizers of cell structure and function.Crossref | GoogleScholarGoogle Scholar | 21169733PubMed |

Ohki-Hamazaki, H. (2000). Neuromedin B. Prog. Neurobiol. 62, 297–312.
Neuromedin B.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXlvFymtL4%3D&md5=f2f30db160dc3161ab9d909dcdf92a35CAS | 10840151PubMed |

Olson, E. N., and Nordheim, A. (2010). Linking actin dynamics and gene transcription to drive cellular motile functions. Nat. Rev. Mol. Cell Biol. 11, 353–365.
Linking actin dynamics and gene transcription to drive cellular motile functions.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXltV2lurs%3D&md5=4845fb87ffac875274e50c38389e0dbdCAS | 20414257PubMed |

Padua, M. B., and Hansen, P. J. (2010). Evolution and function of the uterine serpins (SERPINA14). Am. J. Reprod. Immunol. 64, 265–274.
Evolution and function of the uterine serpins (SERPINA14).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhtlSgtrbF&md5=4ba84c3cc7dca1778d89b35e03688dedCAS | 20678169PubMed |

Picard-Tools (2013) A set of tools for working with next generation sequencing data in the BAM format. Available at: http://broadinstitute.github.io/picard/ [verified 24 July 2015].

Pryce, J. E., Nielsen, B. L., Veerkamp, R. F., and Simm, G. (1999). Genotype and feeding system effects and interactions for health and fertility traits in dairy cattle. Livest. Prod. Sci. 57, 193–201.
Genotype and feeding system effects and interactions for health and fertility traits in dairy cattle.Crossref | GoogleScholarGoogle Scholar |

R Development Core Team (2012). ‘R: A Language and Environment for Statistical Computing (2.15 ed.).’ Available at http://web.mit.edu/r_v3.0.1/fullrefman.pdf [Verified 24 July 2015]

Reveille, J. D. (2006). The genetic basis of autoantibody production. Autoimmun. Rev. 5, 389–398.
The genetic basis of autoantibody production.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XhtVGiu7rP&md5=ab2348cfaa7653818ff6a29d6bcc65d0CAS | 16890892PubMed |

Rizos, D., Carter, F., Besenfelder, U., Havlicek, V., and Lonergan, P. (2010). Contribution of the female reproductive tract to low fertility in postpartum lactating dairy cows. J. Dairy Sci. 93, 1022–1029.
Contribution of the female reproductive tract to low fertility in postpartum lactating dairy cows.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXitlOjurc%3D&md5=b3637df12efaa2f7d45f6f0f537bd0b0CAS | 20172222PubMed |

Robinson, M. D., and Oshlack, A. (2010). A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biol. 11, R25.
A scaling normalization method for differential expression analysis of RNA-seq data.Crossref | GoogleScholarGoogle Scholar | 20196867PubMed |

Robinson, M. D., McCarthy, D. J., and Smyth, G. K. (2010). edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26, 139–140.
edgeR: a Bioconductor package for differential expression analysis of digital gene expression data.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhs1WlurvO&md5=6c4f2e1ff0fffb45078ce114defde940CAS | 19910308PubMed |

Salilew-Wondim, D., Holker, M., Rings, F., Ghanem, N., Ulas-Cinar, M., Peippo, J., Tholen, E., Looft, C., Schellander, K., and Tesfaye, D. (2010). Bovine pretransfer endometrium and embryo transcriptome fingerprints as predictors of pregnancy success after embryo transfer. Physiol. Genomics 42, 201–218.
Bovine pretransfer endometrium and embryo transcriptome fingerprints as predictors of pregnancy success after embryo transfer.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhtleqsrzN&md5=6ba9d9d40a04b8f05ed5846ff9ef97f5CAS | 20388838PubMed |

Shimizu, T., Krebs, S., Bauersachs, S., Blum, H., Wolf, E., and Miyamoto, A. (2010). Actions and interactions of progesterone and estrogen on transcriptome profiles of the bovine endometrium. Physiol. Genomics 42A, 290–300.
Actions and interactions of progesterone and estrogen on transcriptome profiles of the bovine endometrium.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXmslWgsbo%3D&md5=e88338a1bdf4c82eae858c357d40bff0CAS | 20876846PubMed |

Ulbrich, S. E., Groebner, A. E., and Bauersachs, S. (2013). Transcriptional profiling to address molecular determinants of endometrial receptivity: lessons from studies in livestock species. Methods 59, 108–115.
Transcriptional profiling to address molecular determinants of endometrial receptivity: lessons from studies in livestock species.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhvVWgtbnL&md5=d4d69d5fce83f438c7e9a9290b6b4427CAS | 23178633PubMed |

Veerkamp, R. F., Dillon, P., Kelly, E., Cromie, A. R., and Groen, A. F. (2002). Dairy cattle breeding objectives combining yield, survival and calving interval for pasture-based systems in Ireland under different milk quota scenarios. Livest. Prod. Sci. 76, 137–151.

Velazquez, M. A., Spicer, L. J., and Wathes, D. C. (2008). The role of endocrine insulin-like growth factor-I (IGF-I) in female bovine reproduction. Domest. Anim. Endocrinol. 35, 325–342.
The role of endocrine insulin-like growth factor-I (IGF-I) in female bovine reproduction.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXht1ejt7nL&md5=d7034205171ad4a844890912174c1545CAS | 18703307PubMed |