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Investigation of conceptus stimulated gene expression in buffalo peripheral blood mononuclear cells as potential diagnostic markers of early pregnancy

Published online by Cambridge University Press:  29 May 2023

Paramjeet Sharma ,
Ratan Kumar Choudhary ,
Navdeep Singh Ratta  and
Sikh Tejinder Singh
Paramjeet Sharma
Affiliation:
Animal Stem Cells Lab, Department of Bioinformatics, College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, India
Ratan Kumar Choudhary*
Affiliation:
Animal Stem Cells Lab, Department of Bioinformatics, College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, India
Navdeep Singh Ratta
Affiliation:
Directorate of Livestock Farms, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, India
Sikh Tejinder Singh
Affiliation:
Directorate of Livestock Farms, College of Veterinary Sciences, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, India
*
Corresponding author: Ratan Kumar Choudhary; Email: vetdrrkc@gmail.com
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Abstract

Exploration of novel strategies for early pregnancy diagnosis is pivotal in enhancing the reproductive potential and monetary gains from dairy herds. In buffalo, the trophectoderm cells of the elongating conceptus secrete interferon-tau that stimulates the transcription of various genes in peripheral blood mononuclear cells (PBMC) during the peri-implantation period. We explored the differential expression of classical (ISG15) and novel (LGALS3BP and CD9) early pregnancy markers in PBMC of buffaloes during various stages of pregnancy. Natural heat was detected in buffaloes by assessing the vaginal fluid, and artificial insemination (AI) was done. Whole blood was collected from the jugular vein in EDTA-containing vacutainers for PBMC isolation before AI (0-day) and 20, 25 and 40 d post-AI. On day 40, transrectal ultrasonography examination was performed to confirm pregnancy. The inseminated non-pregnant animals served as control. Total RNA was extracted using the TRIzol method. The temporal abundance of ISG15, LGALS3BP and CD9 genes in PBMC was compared between pregnant and non-pregnant groups (n = 9 per group) using real time-qPCR. Results showed transcripts of ISG15 and LGALS3BP were more abundant at 20 d in the pregnant group compared to the 0 d and 20 d values of the non-pregnant group. However, due to variability in expression, threshold (Ct) cycle of RT-qPCR alone could not distinguish pregnant and non-pregnant animals. In conclusion, ISG15 and LGALS3BP transcripts abundance in PBMCs are potential candidate biomarkers for early prediction of buffalo pregnancy 20-days post-AI, but further work is required to allow the development of a reliable new methodology.

Keywords

Early pregnancyISG15LGALS3BPRT-qPCRultrasound
Type
Research Article
Information
Journal of Dairy Research , Volume 90 , Issue 2 , May 2023 , pp. 142 - 145
Copyright
Copyright © The Author(s), 2023. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

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References

Baba, NA, Panigrahi, M, Verma, AD, Sadam, A, Sulabh, S, Chhotaray, S, Parida, S, Krishnaswamy, N and Bhushan, B (2019) Endometrial transcript profile of progesterone-regulated genes during early pregnancy of Water Buffalo (Bubalus bubalis). Reproduction in Domestic Animals 54, 100–107.Google Scholar
Batra, K, Kumar, A, Maan, S, Kumar, V, Kumari, R and Nanda, T (2018) Recombinant interferon stimulated protein 15 (rISG15) as a molecular marker for detection of early pregnancy in Bubalus bubalis. Animal Reproduction Science 2018(197), 106–116.CrossRefGoogle Scholar
Bauersachs, S, Ulbirch, SE, Gross, K, Schmidt, SEM, Meyer, HHD, Wenigerkind, H, Vermehren, M, Sinowatz, F, Blum, H and Wolf, E (2006) Embryo-induced transcriptome changes in bovine endometrium reveal species-specific and common molecular markers of uterine receptivity. Reproduction (Cambridge, England) 132, 319–331.CrossRefGoogle ScholarPubMed
Buragohain, L, Kumar, R, Nanda, T, Phulia, SK, Mohanty, AK, Kumar, S, Balhara, S, Ghuman, SPS, Singh, I and Balhara, AK (2016) Serum MX2 protein as candidate biomarker for early pregnancy diagnosis in buffalo. Reproduction in Domestic Animals 51(4), 453–60.CrossRefGoogle ScholarPubMed
Casano, AB, Barile, VL, Menchetti, L, Guelfi, G, Brecchia, G, Agradi, S, De Matteis, G, Scatà, MC, Grandoni, F and Barbato, O (2022) Interferon tau (IFNt) and interferon-stimulated genes (ISGs) expression in peripheral blood leukocytes and correlation with circulating pregnancy-associated glycoproteins (PAGs) during peri-implantation and early pregnancy in buffalo cows. Animals (Basel) 12(22), 3068.Google Scholar
Department of Animal Husbandry and dairying (2019) 20th Livestock Census. Available at https://www.dahd.nic.in/sites/default/filess/Key%20Results%2BAnnexure%2018.10.2019.pdf.Google Scholar
Ferraz, PA, Filho, CASG, Rocha, CC, Neto, AL, de Andrade Bruni, G, Oshiro, TSI, Baruselli, PS, Lima, FS and Pugliesi, G (2021) Feasibility and accuracy of using different methods to detect pregnancy by conceptus-stimulated genes in dairy cattle. JDS Communications 2, 153–158.CrossRefGoogle ScholarPubMed
Livak, KJ and Schmittgen, TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods (San Diego, Calif.) 25, 402–408.Google Scholar
Pugliesi, G, Miagawa, BT, Paiva, YN, França, MR, Silva, LA and Binelli, M (2014) Conceptus-induced changes in the gene expression of blood immune cells and the ultrasound-accessed luteal function in beef cattle: how early can we detect pregnancy? Biology of Reproduction 91, 95.CrossRefGoogle ScholarPubMed
Rocha, CC, da Silva Andrade, SC, de Melo, GD, Motta, IG, Coutinho, LL, Gonella-Diaza, AM, Binelli, M and Pugliesi, G (2020) Early pregnancy-induced transcripts in peripheral blood immune cells in Bos indicus heifers. Scientific Reports 10, 1–15.Google Scholar
SAS Institute Inc (2011) SAS 9.3. Help and Documentation. Cary, NC: SAS Institute Inc.Google Scholar
Selvam, RM and Archunan, G (2017) A combinatorial model for effective estrus detection in Murrah buffalo. Veterinary World 10, 209–213.CrossRefGoogle ScholarPubMed
Soumya, NP, Das, DN, Jeyakumar, S, Mondal, S, Mor, A and Mundhe, UT (2017) Differential expression of ISG15 mRNA in peripheral blood mononuclear cells of nulliparous and multiparous pregnant vs. non-pregnant Bos indicus cattle. Reproduction in Domestic Animals 52, 97–106.CrossRefGoogle Scholar
Weimin, L, Yujing, C, Jing, L, Zeng, H and Ping, Z (2007) The expression of CD9 in the peri-implantation mouse uterus is upregulated in an ovarian steroid hormone – dependent manner. Fertility and Sterility 87, 664–670.CrossRefGoogle Scholar
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