Using real-time PCR to identify pregnancy-associated glycoprotein 2 (PAG-2) in water buffalo (Bubalus bubalis) blood in early pregnancy

Barbato, O.; Guelfi, G.; Barile, V. L.; Menchetti, L.; Tortiello, C.; Canali, C.; Brecchia, G.; Traina, G.; Beckers, Jean-François; Melo de Sousa, Noelita

doi:10.1016/j.theriogenology.2016.09.038

Article (Scientific journals)

Using real-time PCR to identify pregnancy-associated glycoprotein 2 (PAG-2) in water buffalo (Bubalus bubalis) blood in early pregnancy

Barbato, O.; Guelfi, G.; Barile, V. L. et al.

2017 • In Theriogenology, 89, p. 106-113

Peer Reviewed verified by ORBi

Permalink
https://hdl.handle.net/2268/250855

DOI
10.1016/j.theriogenology.2016.09.038

PubMed
28043340

Files (1)Send to Details Statistics Bibliography Similar publications

Files

Full Text

1-s2.0-S0093691X16304575-main.pdf

Publisher postprint (765.74 kB)

Request a copy

All documents in ORBi are protected by a user license.

Send to

RIS BibTex APA Chicago Permalink X Linkedin

Details

Keywords :

Polymerase chain reaction; Pregnancy; Pregnancy-associated glycoprotein 2; Water buffalo; Animals; Aspartic Acid Endopeptidases; Buffaloes; Estrus Synchronization; Female; Insemination, Artificial; Placenta; Pregnancy Tests; Pregnancy, Animal; Real-Time Polymerase Chain Reaction; RNA, Messenger

Abstract :

[en] This study investigates for the first time mRNA pregnancy-associated glycoprotein 2 (PAG-2) expression in blood cells during early pregnancy in water buffalo. The PAGs constitute a large family of glycoproteins expressed in the outer epithelial layer of the placenta in eutherian species. All PAGs are not concomitantly expressed throughout pregnancy; some of them are expressed in the earlier phases, whereas others appear later and are expressed over a shorter period. Twenty-one lactating buffaloes were analyzed—17 females were synchronized with PRID and artificially inseminated (AI), whereas four females were synchronized but not inseminated (control group). Blood was collected at Days 0, 18, 28, 40, and 75 from AI (AI = Day 0). Expression of PAG-2 mRNA in blood samples was measured with real-time polymerase chain reaction. Pregnancy diagnosis was performed on Day 28 (D28) and Day 40 (D40) after AI by ultrasonography (US) and by PAG-1 RIA method. The females diagnosed pregnant at D28 and confirmed pregnant at D40 were defined as D28(+)D40(+) group; the females diagnosed pregnant at D28 but not confirmed pregnant at D40 were defined as D28(+)D40(−) group; and the females that were diagnosed as nonpregnant on either days were defined as D28(−)D40(−) group. PAG-2 mRNA at Day 0 was not observed in any groups. The D28(+)D40(+) group showed the highest expression, starting on Day 18 and increasing progressively up to Day 75. PAG-2 mRNA was also expressed on Day 18 in both D28(+)D40(−) and D28(−)D40(−) groups, but their levels were lower than those of D28(+)D40(+) group and almost constant over time. PAG-2 mRNA was never detected in the control group. The significant difference in the expression of PAG-2 mRNA between the D28(+)D40(+) group and the D28(−)D40(−) group, starting from Day 18, suggests that these animals might have conceived, but have experienced early embryonic loss; therefore, the PAG-2 mRNA was still present in blood circulation although at lower levels, as found in the D28(+)D40(−) group. In conclusion, this study shows that PAG-2 mRNA can be detected in peripheral maternal blood cells earlier than circulating PAG-1 molecules and could be useful for studies on early pregnancy and embryonic mortality. © 2016 Elsevier Inc.

Disciplines :

Veterinary medicine & animal health

Author, co-author :

Barbato, O.; Department of Veterinary Medicine, University of Perugia, Perugia, Italy

Guelfi, G.; Department of Veterinary Medicine, University of Perugia, Perugia, Italy

Barile, V. L.; Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria (CREA) – Animal Production Research Centre, Monterotondo, Rome, Italy

Menchetti, L.; Department of Veterinary Medicine, University of Perugia, Perugia, Italy

Tortiello, C.; Department of Veterinary Medicine, University of Perugia, Perugia, Italy

Canali, C.; Department of Veterinary Medicine, University of Perugia, Perugia, Italy

Brecchia, G.; Department of Veterinary Medicine, University of Perugia, Perugia, Italy

Traina, G.; Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy

Beckers, Jean-François ; Université de Liège - ULiège > Département des sciences fonctionnelles (DSF) > Département des sciences fonctionnelles (DSF)

Melo de Sousa, Noelita ; Université de Liège - ULg

Language :

English

Title :

Using real-time PCR to identify pregnancy-associated glycoprotein 2 (PAG-2) in water buffalo (Bubalus bubalis) blood in early pregnancy

Publication date :

2017

Journal title :

Theriogenology

ISSN :

0093-691X

eISSN :

1879-3231

Publisher :

Elsevier, Netherlands

Volume :

Pages :

106-113

Peer reviewed :

Peer Reviewed verified by ORBi

Funders :

Fondazione Cassa Risparmio Perugia [IT]

Available on ORBi :

since 12 September 2020

Statistics

Number of views

61 (1 by ULiège)

Number of downloads

1 (1 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

[1] Oltenacu, P.A., Ferguson, J.D., Lednor, A.J., Economic evaluation of pregnancy diagnosis in dairy cattle: a decision analysis approach. J Dairy Sci 73 (1990), 2826–2831.
[2] Romano, J.E., Thompson, J.A., Kraemer, D.C., Westhusin, M.E., Tomaszweski, M.A., Forrest, D.W., Effects of early pregnancy diagnosis by palpation per rectum on pregnancy loss in dairy cattle. J Am Vet Med Assoc 239 (2011), 668–673.
[3] Diskin, M.G., Parr, M.H., Morris, D.G., Embryo death in cattle: an update. Reprod Fertil Dev 24 (2011), 244–251.
[4] Fricke, P.M., Scanning the future-ultrasonography as a reproductive management tool for dairy cattle. J Dairy Sci 85 (2002), 1918–1926.
[5] Barile, V.L., Improving reproductive efficiency in female buffaloes. Liv Prod Sci 92 (2005), 183–194.
[6] Barile, V.L., Terzano, G.M., Pacelli, C., Todini, L., Malfatti, A., Barbato, O., LH peak and ovulation after two different estrus synchronization treatments in buffalo cows in the daylight-lengthening period. Theriogenology 84 (2015), 286–293.
[7] Sousa, N.M., Beckers, J.F., Gajewski, Z., Current trends in follow-up of trophoblastic function in ruminant species. J Physiol Pharmacol 59 (2008), 65–74.
[8] Taverne, M.A., Szenci, O., Szétag, J., Piros, A., Pregnancy diagnosis in cows with linear-array real-time ultrasound scanning: a preliminary note. Vet Q 7 (1985), 264–270.
[9] Kastelic, J.P., Bergfelt, D.R., Ginther, O.J., Ultrasonic detection of the conceptus and characterization of intrauterine fluid on days 10 to 22 in heifers. Theriogenology 35 (1991), 569–581.
[10] Ginther, O.J., Nuti, L., Wentworth, B.C., Tyler, W.J., Progesterone concentration in milk and blood during pregnancy in cows. Proc Soc Exp Biol Med 146 (1974), 354–357.
[11] Sasser, R.G., Ruder, C.A., Ivani, K.A., Butler, J.E., Hamilton, W.C., Detection of pregnancy by radioimmunoassay of a novel pregnancy-specific protein in serum of cows and a profile of serum concentrations during gestation. Biol Reprod 35 (1986), 936–942.
[12] Zoli, A.P., Guibault, L.A., Delahaut, P., Benitez Ortiz, W., Beckers, J.F., Radioimmunoassay of a bovine pregnancy-associated glycoprotein in serum: its application for pregnancy diagnosis. Biol Reprod 46 (1992), 83–92.
[13] Szenci, O., Beckers, J.F., Sulon, J., Sasser, G., Taverne, M.A.M., Varga, J., et al. Comparison of ultrasonography, bovine pregnancy specific B, and bovine pregnancy-associated glycoprotein 1 test for pregnancy detection in dairy cows. Theriogenology 50 (1998), 77–88.
[14] Breukelman, S.P., Perenyi, Z., Ruigh, L., van Wagtendonk-de Leeuw, A.M., Jonker, F.H., Vernooij, J.C., et al. Plasma concentrations of bovine pregnancy-associated glycoprotein (bPAG) do not differ during the first 119 days between ongoing pregnancies derived by transfer of in vivo and in vitro produced embryos. Theriogenology 63 (2005), 1378–1389.
[15] Forde, N., Lonergan, P., Transcriptomic analysis of the bovine endometrium: what is required to establish uterine receptivity to implantation in cattle?. J Reprod Dev 58 (2012), 189–195.
[16] Han, H., Austin, K.J., Rempel, L.A., Hansen, T.R., Low blood ISG15 MRNA and progesterone levels are predictive of non-pregnant dairy cows. J Endocrinol 191 (2006), 505–512.
[17] Stevenson, J.L., Dalton, J.C., Ott, T.L., Racicot, K.E., Chebel, R.C., 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 85 (2007), 2163–2172.
[18] Green, J.C., Okamura, C.S., Poock, S.E., Lucy, M.C., Measurement of interferon-tau (IFN-tau) stimulated gene expression in blood leukocytes for pregnancy diagnosis within 18-20 d after insemination in dairy cattle. Anim Reprod Sci 121 (2010), 24–33.
[19] Barbato, O., Merlo, M., Celi, P., Sousa, N.M., Guarneri, L., Beckers, J.F., et al. Relationship between plasma progesterone and pregnancy-associated glycoprotein concentrations during early pregnancy in dairy cows. Vet J 195 (2013), 385–387.
[20] Hughes, A.L., Green, J.A., Garbayo, J.M., Roberts, R.M., Adaptive diversification within a large family of recently duplicated, placentally expressed genes. Proc Natl Acad Sci U S A 97 (2000), 3319–3323.
[21] Green, J.A., Xie, S., Quan, X., Bao, B., Gan, X., Mathialogan, N., et al. Pregnancy-associated bovine and ovine glycoproteins exhibit spatially and temporally distinct expression patterns during pregnancy. Biol Reprod 62 (2000), 1624–1631.
[22] Xie, S., Low, B.G., Nagel, R.J., Beckers, J.F., Roberts, R.M., A novel glycoprotein of the aspartic proteinase gene family expressed in bovine placental trophectoderm. Biol Reprod 51 (1994), 1145–1153.
[23] Garbayo, J.M., Green, J., Manikkam, M., Beckers, J.F., Kiesling, D., Ealy, A.D., et al. Caprine pregnancy-associated glycoproteins (PAG): their cloning, expression and evolutionary relationship to other PAG. Mol Reprod Dev 57 (2000), 311–322.
[24] Barbato, O., Sousa, N.M., Klisch, K., Clerget, E., Debenedetti, A., Barile, V.L., et al. Isolation of new pregnancy-associated glycoproteins from water buffalo (Bubalus bubalis) placenta by Vicia villosa affinity chromatography. Res Vet Sci 85 (2008), 457–466.
[25] Barbato, O., Melo de Sousa, N., Barile, V.L., Canali, C., Beckers, J.F., Purification of pregnancy-associated glycoproteins from late-pregnancy Bubalus bubalis placentas and development of a radioimmunoassay for pregnancy diagnosis in water buffalo females. BMC Vet Res, 9, 2013, 89.
[26] Sharma, R.K., Singh, J.K., KHanna, S., Phulia, S.K., Sarkar, S.K., Singh, I., Fetal age determination in Murrah buffaloes from days 22 through 60 with ultrasonography. Indian J Anim Sci 82 (2012), 374–376.
[27] Zoli, A.P., Beckers, J.F., Wouters-Ballman, P., Closset, J., Falmagne, P., Ectors, F., Purification and characterization of a bovine pregnancy-associated glycoprotein. Biol Reprod 45 (1991), 1–10.
[28] Diverio, S., Guelfi, G., Barbato, O., Di Mari, W., Egidi, G., Santoro, M.M., Non-invasive assessment of animal exercise stress: real-time PCR of GLUT4, COX2, SOD1 and HSP70 in avalanche military dog saliva. Animal 5 (2015), 774–792.
[29] López-Gatius, F., Hunter, R.H., Garbayo, J.M., Santolaria, P., Yániz, J., Serrano, B., et al. Plasma concentrations of pregnancy-associated glycoprotein-1 (PAG-1) in high producing dairy cows suffering early fetal loss during the warm season. Theriogenology 67 (2007), 1324–1330.
[30] Nguyen, V.H., Barbato, O., Bui, X.N., Beckers, J.F., Sousa, N.M., Assessment of pregnancy-associated glycoprotein (PAG) concentrations in swamp buffalo samples from fetal and maternal origins by using interspecies antisera. Anim Sci J 83 (2012), 683–689.
[31] Breukelman, S.P., Perényi, Z., Taverne, M.A.M., Jonker, H., van der Weijden, G.C., Vos, P.L.A., et al. Characterisation of pregnancy losses by measuring plasma profiles of progesterone and bovine pregnancy-associated glycoprotein-1 after embryo transfer. Vet J 194 (2012), 71–76.
[32] Barbato, O., Sousa, N.M., Debenedetti, A., Canali, C., Todini, L., Beckers, J.F., Validation of a new pregnancy-associated glycoprotein radioimmunoassay method for the detection of early pregnancy in ewes. Theriogenology 72 (2009), 993–1000.
[33] Karen, A., Darwish, S., Ramoun, A., Tawfeek, K., Van Han, N., Sousa, N.M., et al. Accuracy of ultrasonography and pregnancy-associated glycoprotein test for pregnancy diagnosis in buffaloes. Theriogenology 68 (2007), 1150–1155.
[34] Barbato, O., Chiardia, E., Barile, V.L., Pierri, F., Melo de Sousa, N., Terracina, L., et al. Investigation into omocysteine, vitamin E and malondialdeyde as indicators of successful artificial insemination in synchronized buffalo cows (Bubalus bubalis). Res Vet Sci 104 (2016), 100–105.
[35] Gifford, C.A., Racicot, K., Clark, D.S., Austin, K.J., Hansen, T.R., Lucy, M.C., et al. Regulation of interferon-stimulated genes in peripheral blood leukocytes in pregnant and bred, non pregnant dairy cows. J Dairy Sci 90 (2007), 274–280.
[36] Roberts, R.M., Cross, J.C., Leaman, D.W., Interferons as hormones of pregnancy. Endocr Rev 13 (1992), 432–452.
[37] Spencer, T.E., Bazer, F.W., Biology of progesterone action during pregnancy recognition and maintenance of pregnancy. Front Biosci 7 (2002), 1879–1898.
[38] Chethan, S.G., Singh, S.K., Nongsiej, J., Rakesh, H.B., Singh, R.P., Kumar, N., et al. IFN-τ acts in a dose-dependent manner on prostaglandin production by buffalo endometrial stromal cells cultured in vitro. Reprod Domest Anim 49 (2014), 403–408.
[39] Sandra, O., Constanta, C., Carvalho, A.V., Eozénoua, C., Valoura, D., Mauffréa, V., et al. Maternal organism and embryo biosensoring: insights from ruminants. J Reprod Immunol 108 (2015), 105–113.
[40] Ushizawa, K., Herath, C.B., Kaneyama, K., Shiojima, S., Hirasawa, A., Takahashi, T., et al. cDNA microarray analysis of bovine embryo gene expression profiles during the pre-implantation period. Reprod Biol Endocrinol, 2, 2004, 77.
[41] Mamo, S., Mehta, J.P., McGettigan, P., Fair, T., Spencer, T.E., Bazer, F.W., et al. RNA sequencing reveals novel gene clusters in bovine conceptuses associated with maternal recognition of pregnancy and implantation. Biol Reprod 85 (2011), 1143–1151.
[42] Thompson, I.M., Cerri, R.L., Kim, I.H., Ealy, A.D., Hansen, P.J., Staples, C.R., et al. Effects of lactation and pregnancy on metabolic and hormonal responses and expression of selected conceptus and endometrial genes of Holstein dairy cattle. J Dairy Sci 95 (2012), 5645–5656.
[43] Hue, I., Degrelle, S.A., Turenne, N., Conceptus elongation in cattle. Genes, models and questions. Anim Reprod Sci 134 (2012), 19–28.
[44] Del Vecchio, R.P., Sutherland, W.D., Sasser, R.G., Bovine luteal cell production in vitro of prostaglandin E2, oxytocin and progesterone in response to pregnancy-specific protein B and prostaglandin F2α. J Reprod Fertil 107 (1996), 131–136.
[45] Weems, S.Y., Lammoglia, M.A., Vera-Avila, H.R., Randel, R.D., King, C., Sasser, R.G., et al. Effect of luteinizing hormone (LH), PGE2, 8-EPI-PGE1, 8-EPI-PGE2, trichosanthin, and pregnancy specific protein B (PSPB) on secretion of progesterone in vitro by corpora lutea (CL) from nonpregnant and pregnant cows. Prostaglandins Other Lipid Mediat 55 (1998), 27–42.
[46] Austin, K.J., King, L.P., Vierk, J.E., Spencer, R.C., Hauren, T.R., Pregnancy-specific-protein B induces release on an alpha chemokine in bovine endometrium. Endocrinology 140 (1999), 542–545.
[47] Hoeben, D., Burvenicha, C., Massart-Leena, A., Lenjou, M., Nij, G., In vitro effect of ketone bodies, glucocorticosteroids and bovine pregnancy-associated glycoprotein on cultures of bone marrow progenitor cells of cows and calves. Vet Immunol Immunopathol 68 (1999), 229–240.