Animal Reproduction (AR)
Animal Reproduction (AR)

Human Chorionic Gonadotrophin (hCG) induces changes in IFN-pathway and Interferon-Stimulated Genes (ISGs) on the bovine endometrium at Day 18 of pregnancy

Manuela Wolker Manta; Eduardo Pradebon da Silva; Suzana Rossato Feltrin; Amanda Luiza Prante; Karine de Vargas Aires; Leonardo Guedes de Andrade; Ana Paula da Silva; Carolina dos Santos Amaral; Letícia Minussi Wink; Valério Marques Portela; Alfredo Quites Antoniazzi

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Abstract: We hypothesized that the hCG modulates the expression of IFNT-pathway and ISGs in bovine endometrium during early pregnancy. The aim of the current study is to evaluate the effect of hCG on IFNT-pathway signals and ISGs expression in endometrial cells. For this, 29 non-lactating cross-bread cows were used in the study and submitted to a 9-day fixed-time artificial insemination (FTAI) protocol. The day of the AI was considered Day 0 (D0), and five days (D5) after the FTAI, the cows were allocated into two groups: Control and hCG group, when a hCG group received a single dose of 2.500UI of hCG. On day 18 after FTAI (D18) cows were slaughtered and endometrial tissue samples were collected. There was no difference between the embryo recovery rate of the cows in C compared to the hCG. The hCG group increased the accessory corpus luteum formation rate. The hCG resulted in greater serum progesterone concentration in the hCG group compared to the C on Day 14. Only the expression of IFNAR2 and STAT1 were upregulated on pregnant cows of the hCG group compared to the C group. The pathway genes (JAK1, STAT2, and IRF9) were not regulated. The mRNA abundance of ISG15, MX1, MX2, and OAS1 was upregulated in pregnant cows for hCG group, compared to C group. The results show that the administration of hCG, 5 days after AI, in addition to increasing the serum progesterone, modulates the expression of IFNT-pathway and ISGs on bovine endometrium on Day 18 of pregnancy.


IFNT, hCG, pregnancy, IFN-pathway, ISGs


Amaral CD, Correa GRE, Serrano Mujica LK, Fiorenza MF, Rosa SG, Nogueira CW, Portela VM, Comim FV, Schoenau W, Smirnova NP, Antoniazzi AQ. Heat stress modulates polymorphonuclear cell response in early pregnancy cows: I. interferon pathway and oxidative stress. PLoS One. 2021;16(9):e0257418. PMid:34543299.

Austin KJ, Ward SK, Teixeira MG, Dean VC, Moore DW, Hansen TR. Ubiquitin cross-reactive protein is released by the bovine uterus in response to interferon during early pregnancy. Biol Reprod. 1996;54(3):600-6. PMid:8835381.

Bazer FW. Establishment of pregnancy in sheep and pigs. Reprod Fertil Dev. 1989;1(3):237-42. PMid:2682807.

Bazer FW, Kim J, Song G, Ka H, Tekwe CD, Wu G. Select nutrients, progesterone, and interferon tau affect conceptus metabolism and development. Ann N Y Acad Sci. 2012;1271(1):88-96. PMid:23050969.

Beltran MP, Vasconcelos JLM. Conception rate in Holstein cows treated with GnRH or hCG on the fifth day post artificial insemination during summe. Arq Bras Med Vet Zootec. 2008;60(3):580-6.

Bettegowda A, Patel OV, Ireland JJ, Smith GW. Quantitative analysis of messenger RNA abundance for ribosomal protein L-15, cyclophilin-A, phosphoglycerokinase, beta-glucuronidase, glyceraldehyde 3-phosphate dehydrogenase, beta-actin, and histone H2A during bovine oocyte maturation and early embryogenesis in vitro. Mol Reprod Dev. 2006;73(3):267-78. PMid:16261607.

Cheng Z, Brown LE, Wathes DC. Bovine Viral diarrhoea virus infection disrupts uterine interferon stimulated gene regulatory pathways during pregnancy recognition in cows. Viruses. 2019;12(1):1. PMid:31861316.

Clemente M, Lopez-Vidriero I, O’Gaora P, Mehta JP, Forde N, Gutierrez-Adan A, Lonergan P, Rizos D. Transcriptome changes at the initiation of elongation in the bovine conceptus. Biol Reprod. 2011;85(2):285-95. PMid:21508349.

David M. Signal transduction by type I interferons. Biotechniques. 2002;33(4S, Suppl):S58 -65. PMid:12395928.

De Rensis F, Lopez-Gatius F, Garcia-Ispierto I, Techakumpu M. Clinical use of human chorionic gonadotropin in dairy cows: an update. Theriogenology. 2010;73(8):1001-8. PMid:20116839.

Edmonson AJ, Lean IJ, Weaver LD, Farver T, Webster G. A body condition scoring chart for Holstein dairy-cows. J Dairy Sci. 1989;72(1):68-78.

Forde N, Carter F, di Francesco S, Mehta JP, Garcia-Herreros M, Gad A, Tesfaye D, Hoelker M, Schellander K, Lonergan P. Endometrial response of beef heifers on day 7 following insemination to supraphysiological concentrations of progesterone associated with superovulation. Physiol Genomics. 2012;44(22):1107-15. PMid:23012394.

Forde N, Mehta JP, McGettigan PA, Mamo S, Bazer FW, Spencer TE, Lonergan P. Alterations in expression of endometrial genes coding for proteins secreted into the uterine lumen during conceptus elongation in cattle. BMC Genomics. 2013;14(1):321. PMid:23663413.

Freidman S, Gurevich M, Shemesh M. Bovine cyclic endometrium contains high-affinity luteinizing hormone/human chorionic gonadotropin binding sites. Biol Reprod. 1995;52(5):1020-6. PMid:7626701.

Funston RN, Lipsey RJ, Geary TW, Roberts AJ. Effect of administration of human chorionic gonadotropin after artificial insemination on concentrations of progesterone and conception rates in beef heifers. J Anim Sci. 2005;83(6):1403-5. PMid:15890818.

Garrett JE, Geisert RD, Zavy MT, Morgan GL. Evidence for maternal regulation of early conceptus growth and development in beef cattle. J Reprod Fertil. 1988;84(2):437-46. PMid:3199361.

Geisert RD, Morgan GL, Short EC Jr, Zavy MT. Endocrine events associated with endometrial function and conceptus development in cattle. Reprod Fertil Dev. 1992;4(3):301-5. PMid:1438962.

Gray CA, Abbey CA, Beremand PD, Choi Y, Farmer JL, Adelson DL, Thomas TL, Bazer FW, Spencer TE. Identification of endometrial genes regulated by early pregnancy, progesterone, and interferon tau in the ovine uterus. Biol Reprod. 2006;74(2):383-94. PMid:16251498.

Hazano K, Haneda S, Kayano M, Miura R, Matsui M. Effects of hCG administration on corpus luteum development and plasma sex steroid hormone concentration in beef heifers differ according to the locational relationships of the original corpus luteum and the first-wave dominant follicle. J Vet Med Sci. 2020;82(8):1219-25. PMid:32581156.

Horisberger MA. Virus-specific effects of recombinant porcine interferon-gamma and the induction of Mx proteins in pig cells. J Interferon Res. 1992;12(6):439-44. PMid:1337754.

Ivashkiv LB, Hu XY. Signaling by STATs. Arthritis Res Ther. 2004;6(4):159-68. PMid:15225360.

Kraus TA, Lau JF, Parisien JP, Horvath CM. A hybrid IRF9-STAT2 protein recapitulates interferon-stimulated gene expression and antiviral response. J Biol Chem. 2003;278(15):13033-8. PMid:12574168.

Lim JK, Langer JA. Cloning and characterization of a bovine alpha-interferon receptor. Biochim Biophys Acta. 1993;1173(3):314-9. PMid:8318540.

Majoros A, Platanitis E, Kernbauer-Holzl E, Rosebrock F, Muller M, Decker T. Canonical and non-canonical aspects of JAK-STAT signaling: lessons from interferons for cytokine responses. Front Immunol. 2017;8:29. PMid:28184222.

Mann GE, Lamming GE, Robinson RS, Wathes DC. The regulation of interferon-tau production and uterine hormone receptors during early pregnancy. J Reprod Fertil Suppl. 1999;54:317-28. PMid:10692864.

Mansouri-Attia N, Aubert J, Reinaud P, Giraud-Delville C, Taghouti G, Galio L, Everts RE, Degrelle S, Richard C, Hue I, Yang X, Tian XC, Lewin HA, Renard JP, Sandra O. Gene expression profiles of bovine caruncular and intercaruncular endometrium at implantation. Physiol Genomics. 2009;39(1):14-27. PMid:19622795.

Niemann H, Sacher B, Elsaesser F. Pregnancy rates relative to recipient plasma progesterone levels on the day of nonsurgical transfer of frozen/thawed bovine embryos. Theriogenology. 1985;23(4):631-9. PMid:16726032.

Nishigai M, Kamomae H, Tanaka T, Kaneda Y. Improvement of pregnancy rate in Japanese Black cows by administration of hCG to recipients of transferred frozen-thawed embryos. Theriogenology. 2002;58(8):1597-606. PMid:12374129.

O’Hara L, Forde N, Kelly AK, Lonergan P. Effect of bovine blastocyst size at embryo transfer on day 7 on conceptus length on day 14: can supplementary progesterone rescue small embryos? Theriogenology. 2014;81(8):1123-8. PMid:24582375.

Pestka S. The interferons: 50 years after their discovery, there is much more to learn. J Biol Chem. 2007;282(28):20047-51. PMid:17502369.

Roberts RM. Interferon-tau and pregnancy. J Interferon Cytokine Res. 1996;16(4):271-3. PMid:9162519.

Roberts RM, Liu LM, Alexenko A. New and atypical families of type I interferons in mammals: comparative functions, structures, and evolutionary relationships. Prog Nucleic Acid Res Mol Biol. 1997;56(56):287-325. PMid:9187057.

Sá MF Fo, Gonella-Diaza AM, Sponchiado M, Mendanha MF, Pugliesi G, Ramos RDS, Andrade S, Gasparin G, Coutinho LL, Goissis MD, Mesquita FS, Baruselli PS, Binelli M. Impact of hormonal modulation at proestrus on ovarian responses and uterine gene expression of suckled anestrous beef cows. J Anim Sci Biotechnol. 2017;8(1):79. PMid:29118976.

Salzberg S, Hyman T, Turm H, Kinar Y, Schwartz Y, Nir U, Lejbkowicz F, Huberman E. Ectopic expression of 2-5A synthetase in myeloid cells induces growth arrest and facilitates the appearance of a myeloid differentiation marker. Cancer Res. 1997;57(13):2732-40. PMid:9205084.

Seegert D, Strehlow I, Klose B, Levy DE, Schindler C, Decker T. A novel interferon-alpha-regulated, DNA-binding protein participates in the regulation of the IFP53/tryptophanyl-tRNA synthetase gene. J Biol Chem. 1994;269(11):8590-5. PMid:8132584.

Spencer TE, Johnson GA, Bazer FW, Burghardt RC, Palmarini M. Pregnancy recognition and conceptus implantation in domestic ruminants: roles of progesterone, interferons and endogenous retroviruses. Reprod Fertil Dev. 2007;19(1):65-78. PMid:17389136.

Spencer TE, Johnson GA, Burghardt RC, Bazer FW. Progesterone and placental hormone actions on the uterus: insights from domestic animals. Biol Reprod. 2004a;71(1):2-10. PMid:14973264.

Spencer TE, Burghardt RC, Johnson GA, Bazer FW. Conceptus signals for establishment and maintenance of pregnancy. Anim Reprod Sci. 2004b;82-83:537-50. PMid:15271478.

Sponchiado M, Gomes NS, Fontes PK, Martins T, del Collado M, Pastore AD, Pugliesi G, Nogueira MFG, Binelli M. Pre-hatching embryo-dependent and -independent programming of endometrial function in cattle. PLoS One. 2017;12(4):e0175954. PMid:28423001.

Stevenson JL, Dalton JC, Ott TL, Racicot KE, Chebel RC. Correlation between reproductive status and steady-state messenger ribonucleic acid levels of the Myxovirus resistance gene, MX2, in peripheral blood leukocytes of dairy heifers. J Anim Sci. 2007;85(9):2163-72. PMid:17431047.

Stevenson JS, Pulley SL. Characteristics and retention of luteal structures, extended postinsemination cycle, progesterone, and pregnancy-specific protein B in serum after human chorionic gonadotropin treatment of dairy cows. J Dairy Sci. 2012;95(8):4396-409. PMid:22818453.

Stewart MD, Johnson GA, Vyhlidal CA, Burghardt RC, Safe SH, Yu-Lee LY, Bazer FW, Spencer TE. Interferon-τ activates multiple signal transducer and activator of transcription proteins and has complex effects on interferon-responsive gene transcription in ovine endometrial epithelial cells. Endocrinology. 2001a;142(1):98-107. PMid:11145571.

Stewart MD, Johnson GA, Bazer FW, Spencer TE. Interferon-τ (IFNτ) regulation of IFN-stimulated gene expression in cell lines lacking specific IFN-signaling components. Endocrinology. 2001b;142(5):1786-94. PMid:11316742.

Thatcher WW, Bazer FW, Sharp DC, Roberts RM. Interrelationships between uterus and conceptus to maintain corpus luteum function in early pregnancy: sheep, cattle, pigs and horses. J Anim Sci. 1986;62(Suppl 2):25-46. PMid:3533874.

Vitorino Carvalho A, Eozenou C, Healey GD, Forde N, Reinaud P, Chebrout M, Gall L, Rodde N, Padilla AL, Delville CG, Leveugle M, Richard C, Sheldon IM, Lonergan P, Jolivet G, Sandra O. Analysis of STAT1 expression and biological activity reveals interferon-tau-dependent STAT1-regulated SOCS genes in the bovine endometrium. Reprod Fertil Dev. 2016;28(4):459-74. PMid:25116692.

Voelz BE, Cline GF, Hart CG, Lemley CO, Larson JE. Effect of 2 corpora lutea on blood perfusion, peripheral progesterone, and hepatic steroid-inactivating enzymes in dairy cattle. J Dairy Sci. 2015;98(1):288-94. PMid:25468695.

Wang X, Chen CL, Wang LJ, Su YZ, Li BY, Xiao LF, Lin ZL, Sheng XH, Qi XL, Ni HM, Guo Y. Specific activation of embryonic IFNAR1 and endometrial IFNAR2 induced by embryonic IFN tau directs normal uterine fate for bovine early implantation. J Reprod Immunol. 2022;153:103677. PMid:35907379.

Wecker F, Thedy DX, Gonsloroski AV, Borges JBS. Effect of administration of eCG or hCG 7 days after FTAI on the development of ovarian structures and pregnancy rates in beef cows. Acta Sci Vet. 2012;40(4):1-8.

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