Histomorphometry and uterine proteomics during the normal reproductive cycle in bitches

Authors

DOI:

https://doi.org/10.33448/rsd-v10i11.19093

Keywords:

Bitches; Breed; Estrous Cycle; Histology; Proteins; Proteome; Uterus.

Abstract

We aimed to evaluate the histomorphometry and proteomic profile of the canine uterus during all stages of the reproductive cycle. Eighteen healthy female dogs had their estrous cycle identified by clinical evaluation, vaginal cytology, and serum progesterone levels, which were allocated to the proestrus (n=5), estrus (n=5), diestrus (n=5), and anestrus (n=3) groups. All were submitted to elective ovariosalpingohysterectomy, and the uteri were collected for histomorphometric measurement (Image J software). For proteomic analysis, fragments of the uterine horns were subjected to protein measurement (Bradford method) and extraction by 2D electrophoresis (PDquest software). The results showed that the diestrus promoted greater values of thickness in the uterine structures (μm): uterine wall (2,223.8±229.8), endometrium (819.7±109.1), and myometrium (1,392.6±294.2). Uterus showed a protein profile with good reproducibility per phase (pI: 3.5–9.0; PM: 24–150 KDa), with 11 spots in all phases. Despite the greatest histomorphometric changes in the diestrus, we observed a greater number of spots in the estrus (253±45), followed by the proestrus (185±21), diestrus (113±39), and anestrus (80±21). This finding showed probable participation of these proteins in the uterine preparation for receiving gametes for fertilization. Our results showed greater uterine thickness in the diestrus, and greater protein secretion in the estrus, contributing to the prospection of identification of proteins responsible for the biological reproduction processes.

References

Agostinis, C., Mangogna, A., Bossi, F., Ricci, G., Kishore, U. & Bulla, R. (2019). Uterine Immunity and Microbiota: A Shifting Paradigm. Frontiers in Immunology, 10(2387), 1-11.

Aplin, J. D., Fazleabas, A. T., Glasser, S. R. & Giudice, L. C. (2008). The Endometrium: Molecular, Cellular and Clinical Perspectives (2nd ed.). Boca Raton: CRC Press.

Camargo, K. S., Aleixo, G. A. S., Penaforte Júnior, M. A., Galeas, G. R., Trajano, S. C., Melo, K. D., Ferreira, M. S. S., Andrade, L. S. S. & Lopes, L. A. (2019). Achados histopatológicos em úteros e ovários de cadelas submetidas à castração eletiva pelas técnicas de ovariectomia ou ovariohisterectomia. Medicina Veterinária (UFRPE), 13(4), 577–582.

Fahiminiya, S., Reynaud, K., Labas, V., Batard, S., Chastant-Maillard, S. & Gérard, N. (2010). Steroid hormones content and proteomic analysis of canine follicular fluid during the preovulatory period. Reproductive Biology and Endocrinology, 8 (132), 1-13.

Faulkner, S., Elia, G., O' Boyle, P., Dunn, M. & Morris, D. (2013). Composition of the bovine uterine proteome is associated with stage of cycle and concentration of systemic progesterone. Proteomics, 13(22), 3333–3353.

Freitas, L. A., Villamil, P. R., Moura, A. A. A. N., Silva, & L. D. M. (2015). Proteoma uterino durante o ciclo reprodutivo e gestação em animais domésticos. Revista Brasileira de Reprodução Animal, 39(4), 375–381.

Galabova, G., Egerbacher, M., Aurich, J. E., Leitner, M. & Walter, I. (2003). Morphological changes of the endometrial epithelium in the bitch during metoestrus and anoestrus. Reproduction in Domestic Animals, 38(5), 415–420.

Gao, H., Wu, G., Spencer, T. E., Johnson, G. A. & Bazer, F. W. (2009). Select nutrients in the ovine uterine lumen. II. Glucose transporters in the uterus and peri-implantation conceptuses. Biology of Reproduction, 80(1), 94–104.

Holst, P. (2019). Canine Reproduction: The Breeder's Guide (3rd ed.). Dogwise Publishing.

Kuleš, J., Horvatić, A., Guillemin, N., Ferreira, R. F., Mischke, R., Mrljak, V., Chadwick, C. C. & David Eckersall, P. (2020). The plasma proteome and the acute phase protein response in canine pyometra. Journal of Proteomics, 223, 103817.

Lee, W. Y., Chai, S. Y., Lee, K. H., Park, H. J., Kim, J. H., Kim, B., Kim, N. H., Jeon, H. S., Kim, I. C., Choi, H. S. & Song, H. (2013). Identification of the DDAH2 protein in pig reproductive tract mucus: a putative oestrus detection marker. Reproduction in Domestic Animals, 48(1), 13–16.

Monteiro, C. M. R., Perri, S. H. V., Carvalho, R. G., da Silva, A. M. & Koivisto, M. B. (2012). Histomorfometria do corno uterino de gatas (Felis catus) submetidas à ovariosalpingohisterectomia. Brazilian Journal of Veterinary Research and Animal, 49(3), 225–231.

Pereira A. S., Shitsuka, D. M., Parreira, F. J. & Sitsuka, R. (2018). Metodologia da pesquisa científica. UFSM.

Praderio, R. G., García Mitacek, M. C., Núñez Favre, R., Rearte, R., De La Sota, R. L. & Stornelli, M. A. (2019). Uterine endometrial cytology, biopsy, bacteriology, and serum C-reactive protein in clinically healthy diestrus bitches. Theriogenology, 131, 153–161.

Ramos, J. L. G., Ramos, C. L. F. G., Cunha, I. C. N., Carvalho, E. C. Q., Shimoda, E. & Luz, M. R. (2015). Análise histomorfométrica do útero na espécie canina do nascimento aos seis meses de idade. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 67(1), 41–48.

Reynaud, K., Saint-Dizier, M., Tahir, M. Z., Havard, T., Harichaux, G., Labas, V., Thoumire, S., Fontbonne, A., Grimard, B. & Chastant-Maillard, S. (2015). Progesterone plays a critical role in canine oocyte maturation and fertilization. Biology of Reproduction, 93(4), 87.

Salinas, P., Miglino, M. A. & Del Sol, M. (2017). Histomorphometrics and quantitative unbiased stereology in canine uteri treated with medroxyprogesterone acetate. Theriogenology, 95, 105–112.

Swegen, A., Grupen, C. G., Gibb, Z., Baker, M. A., de Ruijter-Villani, M., Smith, N. D., Stout, T. A. E. & Aitken, R. J. (2017). From Peptide Masses to Pregnancy Maintenance: A Comprehensive Proteomic Analysis of The Early Equine Embryo Secretome, Blastocoel Fluid, and Capsule. Proteomics, 17(17–18), 1–30.

Valdés, A., Holst, B. S., Lindersson, S. & Ramström, M. (2019). Development of MS-based methods for identification and quantification of proteins altered during early pregnancy in dogs. Journal of Proteomics, 192, 223–232.

Vermeirsch, H., Simoens, P., Hellemans, A., Coryn, M. & Lauwers, H. (2000). Immunohistochemical detection of progesterone receptors in the canine uterus and their relation to sex steroid hormone levels. Theriogenology, 53(3), 773–788.

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Published

22/08/2021

How to Cite

FREITAS, L. A. de; VASCONCELOS, F. R. .; MOURA, A. A. A. N. .; WALLER, S. B. .; COSTA, P. P. C. .; ROSA, B. M. .; RIBEIRO, W. L. C. .; SILVA, L. D. M. da . Histomorphometry and uterine proteomics during the normal reproductive cycle in bitches. Research, Society and Development, [S. l.], v. 10, n. 11, p. e18101119093, 2021. DOI: 10.33448/rsd-v10i11.19093. Disponível em: https://rsdjournal.org/index.php/rsd/article/view/19093. Acesso em: 24 nov. 2024.

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Section

Agrarian and Biological Sciences