Ovarian surface epithelium of hypothyroid newborn and neonatal rats: from PCNA and caspase-3 perspective

Main Article Content

Anita Radovanović
Milica Kovačević Filipović
Ivan Milošević
Tijana Lužajić
Stefan Veličković
Jana Andrejević
Jelena Danilović Luković


Introduction. The ovarian surface epithelium (OSE) undergoes intensive regeneration and remodelling after each ovulation during the whole reproductive period. This process increases the risk of one of the most common ovarian tumors in women and the female dog. Considering the fact that maternal hypothyroidism highly impacts cell proliferation and cell death during folliculogenesis in the early neonatal period, we aimed to analyze its effect on OSE morphology and dynamics.

Materials and Methods. The study was performed on newborn (24-h-old) and neonatal (4-day-old) female rats, a randomized trial between the control and hypothyroid groups, born under controlled circumstances and hypothyroid mothers, respectively. Their ovaries were analyzed histologically and processed to determine the OSE cell height as an average value of four measurement points. Also, the immunopositivity of the proliferating cell nuclear antigen (PCNA) and caspase-3 were assessed semiquantitatively.

Results and Conclusions. No major structural differences of OSE were found between groups within the given ages except for a slight increment of OSE cell height and incompleteness of apical cell membrane with cytoplasmic projections in hypothyroid animals. PCNA immunopositivity of the OSE cells was higher in ovaries of hypothyroid animals of both ages in comparison to the controls. Moreover, only scarce OSE cells were caspase-3 positive in both groups and ages, with no difference in immunopositivity. Our study confirms the impact of hypothyroidism in the early postnatal period on morphology and proliferation rate of OSE cells, with no effect on caspase-3 dependent cell removal, which may serve as a premise for future investigation of potential carcinogenesis, in terms of prevention and treatment of ovarian cancer.


Download data is not yet available.

Article Details

How to Cite
Radovanović, A., Kovačević Filipović, M., Milošević, I., Lužajić, T., Veličković, S., Andrejević, J., & Danilović Luković, J. (2018). Ovarian surface epithelium of hypothyroid newborn and neonatal rats: from PCNA and caspase-3 perspective. Veterinarski Glasnik, 72(2), 80–89. https://doi.org/10.2298/VETGL180320007R
Full research article


Aghajanova L., Lindeberg M., Carlsson I.B., Stavreus-Evers A., Zhang P., Scott J.E., Skjöldebrand-Sparre L. 2009. Receptors for thyroid-stimulating hormone and thyroid hormones in human ovarian tissue. Reproductive Biomedicine Online, 18(3):337-347. https://doi.org/10.1016/S1472-6483(10)60091-0.

Anreder M.B., Freeman S.M., Merogi A., Halabi S., Marrogi A.J. 1999. p53, c-erbB2, and PCNA status in benign, proliferative, and malignant ovarian surface epithelial neoplasms: A study of 75 cases. Archives of Pathology and Laboratory Medicine, 123(4):310-316.

Auersperg N., Woo M.M., Gilks C.B. 2008. The origin of ovarian carcinomas: a developmental view. Gynecologic Oncology, 110(3):452-454. https://doi.org/10.1016/j.ygyno.2008.05.031.

Caric A., Poljicanin A., Tomic S., Vilovic K., Saraga-Babic M., Vukojevic K. 2014. Apoptotic pathways in ovarian surface epithelium of human embryos during embryogenesis and carcinogenesis: close relationship of developmental plasticity and neoplasm. Acta Histochemica, 116(2):304-311. https://doi.org/10.1016/j.acthis.2013.08.005.

Banco, B., Antuofermo, E., Borzacchiello, G., Cossu-Rocca, P. and Grieco, V., 2011. Canine ovarian tumors: an immunohistochemical study with HBME-1 antibody. Journal of veterinary diagnostic investigation, 23(5):.977-981. http://journals.sagepub.com/doi/abs/10.1177/1040638711416848

Danilović Luković J., Korać A., Milošević I., Lužajić T., Milanović Z., Kovačević Filipović M., Radovanović A. 2017. Z-cells and oogonia/oocytes in advanced process of autophagy are dominant altered cells in ovaries of hypothyroid newborn rats. Acta Veterinaria, Beograd, 67(1):92-106. https://doi.org/10.1515/acve-2017-0009.

Danilović Luković J., Korać A., Milošević I., Lužajić T., Puškaš N., Kovačević Filipović M., Radovanović A. 2016. Altered state of primordial follicles in neonatal and early infantile rats due to maternal hypothyroidism: Light and electron microscopy approach, Micron, 90:33-42. https://doi.org/10.1016/j.micron.2016.08.007.

De Felici M., Lobascio A.M., Klinger F.G. 2008. Cell death in fetal oocytes: many players for multiple pathways, Autophagy, 4(2):240-242. https://doi.org/10.4161/auto.5410.

Escobar M.L., Echeverria O.M., Ortiz R., Vazquez-Nin G.H. 2008. Combined apoptosis and autophagy, the process that eliminates the oocytes of atretic follicles in immature rats, Apoptosis, 13(10):1253-1266. https://doi.org/10.1007/s10495-008-0248-z.

Flesken-Nikitin, A., Hwang, C. I., Cheng, C. Y., Michurina, T. V., Enikolopov, G., & Nikitin, A. Y. 2013. Ovarian surface epithelium at the junction area contains a cancer-prone stem cell niche. Nature, 495(7440):241-245. https://www.nature.com/articles/nature11979.

Fong M.Y., Kakar S.S. 2010. The role of cancer stem cells and the side population in epithelial ovarian cancer. Histology and Histopathology, 25(1):113-120. https:// doi.org/10.14670/HH-25.113.

Giles J.R., Olson L. M., Johnson P.A. 2006. Characterization of ovarian surface epithelial cells from the hen: a unique model for ovarian cancer. Experimental Biology and Medicine, 231(11):1718-1725. https://doi.org/10.1177/153537020623101108.

Gondos B. 1975. Surface epithelium of the developing ovary. Possible correlation with ovarian neoplasia. The American Journal of Pathology, 81(2):303-321.

Kurki P., Ogata K., Tan E.M. 1988. Monoclonal antibodies to proliferating cell nuclear antigen (PCNA)/cyclin as probes for proliferating cells by immunofluorescence microscopy and flow cytometry. Journal of Immunological Methods, 109(1):49-59.

Li S. 1994. Relationship between cellular DNA synthesis, PCNA expression and sex steroid hormone receptor status in the developing mouse ovary, uterus and oviduct. Histochemistry and Cell Biology, 102(5):405-413.

López-Sáez J.F., de la Torre C., Pincheira J., Giménez-Martín G. 1998. Cell proliferation and cancer. Histology and Histopathology, 13(4):1197-1214.

Matikainen T., Perez G.I., Zheng T.S., Kluzak T.R., Rueda B.R., Flavell R.A., Tilly J.L. 2001. Caspase-3 gene knockout defines cell lineage specificity for programmed cell death signaling in the ovary. Endocrinology, 142(6):2468-2480. https://doi.org/10.1210/endo.142.6.8078.

Mostov K., Apodaca G., Aroeti B., Okamoto C. 1992. Plasma membrane protein sorting in polarized epithelial cells. The Journal of Cell Biology, 116(3):577-583.

Murdoch W.J. 2000. Proteolytic and cellular death mechanisms in ovulatory ovarian rupture. Neurosignals, 9(2):102-114. https://doi.org/10.1159/000014629.

Nicholson J.L., Altman J. 1972. The effects of early hypo- and hyperthyroidism on the development of rat cerebellar cortex. I. Cell proliferation and differentiation. Brain Research, 44(1):13-23.

Papadak L., Beilby J.O.W. 1971. The fine structure of the surface epithelium of the human ovary. Journal of Cell Science, 8(2):445-465.

Radovanović A, Roksandić D, Šimić M, Marković D, Gledić D. 2012. Effects of induced maternal hypothyroidism on the ovarian development of offspring rats. Acta veterinaria 62(5-6):483-93. http://www.doiserbia.nb.rs/img/doi/0567-8315/2012/0567-83151206483R.pdf

Rajah R., Glaser E.M., Hirshfield A.N. 1992. The changing architecture of the neonatal rat ovary during histogenesis. Developmental Dynamics, 194(3):177-192.

Talsness C., Grote K., Kuriyama S., Presibella K., Sterner-Kock A., Poça K., Chahoud I. 2015. Prenatal exposure to the phytoestrogen daidzein resulted in persistent changes in ovarian surface epithelial cell height, folliculogenesis, and estrus phase length in adult Sprague-Dawley rat offspring. Journal of Toxicology and Environmental Health, Part A, 78(10):635-644. https://doi.org/10.1080/15287394.2015.1006711.

Tilly J.L., Robles R. 1999. Apoptosis and its impact in clinical reproductive medicine. In Molecular Biology in Reproductive Medicine. Eds. Fauser B.C.J.M., Rutherford A.J., Strauss F.J., Van Steirteghem A., New York, Parthenon, 79–101.

Wu C.T., Altuwaijri S., Ricke W.A., Huang S.P., Yeh S., Zhang C., Chang C. 2007. Increased prostate cell proliferation and loss of cell differentiation in mice lacking prostate epithelial androgen receptor. Proceedings of the National Academy of Sciences, 104(31):12679-12684. https://doi.org/10.1073/pnas.0704940104.

Xu B., Hua J., Zhang Y., Jiang X., Zhang H., Ma T., Zheng W., Sun R., Shen W., Sha J., Cooke H.J., Shi Q. 2011. Proliferating cell nuclear antigen (PCNA) regulates primordial follicle assembly by promoting apoptosis of oocytes in fetal and neonatal mouse ovaries. PLoS One, 6(1):e16046. https://doi.org/10.1371/journal.pone.0016046.

Yazdekhasti H., Rajabi Z., Parvari S., Abbasi M. 2016. Used protocols for isolation and propagation of ovarian stem cells, different cells with different traits. Journal of Ovarian Research, 9(1):68. https://doi.org/10.1186/s13048-016-0274-3.

Most read articles by the same author(s)