Histomorphometry and uterine proteomics during the normal reproductive cycle in bitches
DOI:
https://doi.org/10.33448/rsd-v10i11.19093Keywords:
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.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2021 Luana Azevedo de Freitas; Fábio Roger Vasconcelos; Arlindo Alencar Araripe Noronha Moura; Stefanie Bressan Waller; Paula Priscila Correia Costa; Brenda Madruga Rosa; Wesley Lyeverton Correia Ribeiro; Lúcia Daniel Machado da Silva
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
1) Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
2) Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
3) Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.