221 XENOESTROGENS EXPOSURE AT ENVIRONMENTALLY RELEVANT DOSES ADVERSELY AFFECTS OVINE FOLLICULAR DEVELOPMENT

2010 ◽  
Vol 22 (1) ◽  
pp. 268
Author(s):  
O. E. Rivera ◽  
J. Varayoud ◽  
H. A. Rodríguez ◽  
C. E. Di Mauro ◽  
M. Muãoz-de-Toro ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Androgen Receptor ◽  
Stromal Cells ◽  
Ovarian Development ◽  
Science And Technology ◽  
National Council ◽  
Ki 67 ◽  
Primordial Follicles ◽  
Antral Follicles ◽  
Theca Cells

The exposure to endocrine disrupters may affect reproduction. Our hypothesis suggested that neonatal endocrine disrupter exposure affects ovarian ovine development. Female lambs were s.c. exposed from postnatal Day (PND) 1 to PND 14 to environmentally relevant doses of diethylstilbe- strol (DES; 5 μg kg-1, n = 11), bisphenol A (BPA; 50 μg kg-1, n = 11), or vehicle [control (C), n = 14]. On PND 30, the ovaries were weighed and paraffin-embedded. The whole ovaries were sectioned in sets of 4 adjacent 5-μm serial sections taken 200 μm apart. Follicular dynamics were established by histomorphological features on pricosirius-hematoxylin-stained sections (at least 10 sections/ovary). The total number of fol- licles/ovary was obtained considering the number of sections and section thickness (≥25 000 follicles/ovary). Follicles of different classes and multioocyte follicles were expressed as a percentage. Immunohistochemical expression of estrogen receptor alpha (ERa), androgen receptor (AR), Ki-67 (proliferation marker), and p27 (cyclin-dependent kinase inhibitor 1B) was evaluated. Proliferative index was obtained using Ki-67 expression in the granulose cells (GC) and theca cells of antral follicles. Antral follicles with ≤1% Ki-67 were considered atretic. Kruskal-Wallis and Dunn post-tests were used; P < 0.05 was accepted as significant. In the developing ovaries (PND 1, 5, and 10; n = 3 for each day), the stromal cells showed a high expression of ERa on PND 1, decreasing with age. Estrogen receptor beta (ERfi) was positive in oocytes, CG, and stromal cells, with a temporal pattern opposite to that of ERa. Androgen receptor was found in oocytes, GC, and stromal cells without changes along the studied ages. Results at PND 30 showed that both xenostrogens adversely affect ovarian development. The BPA treatment reduced the ovarian weight without changes in the total number of follicles. Lambs exposed to DES and BPA showed a reduction of the percentage of primordial follicles (C = 59.4 ± 2.9; DES = 36.9 ± 7.3; BPA = 49.8 ± 1.6) but an increase of transitory (C = 29.2 ± 1.8; DES =48.1 ± 4.4; BPA = 41.9 ± 2.5) and pri- mary (C = 1.5 ± 0.2; DES = 5.1 ± 2.0; BPA = 2.9 ± 0.5) follicles. In addition, xenoestrogen treatment increased the number of antral atretic follicles (C = 16.3%; DES = 27.2%; BPA = 49.1%) associated with an increased p27 expression. Besides that, BPA group showed a higher proliferation of GC (C = 12.3 ± 2.6; DES = 31.5 ± 5.3; BPA = 37.0 ± 6.8) and theca cells (C = 1.0 ± 0.3; DES = 7.6 ± 1.9; BPA = 15.1 ± 4.2) in antral follicles and an increased incidence of multioocyte follicles (C = 4.2 ± 1.1; DES = 8.1 ± 2.0; BPA = 18.2 ± 3.9). Our results show that an early exposure to environmentally relevant doses of BPA or DES disrupts the ovine ovarian development and suggest that these effects may be mediated through the early expression of ovarian ER. Xenoestrogen exposure could effect ovarian function with consequences on female fertility. This work was supported by grants from the Universidad Nacional del Litoral (CAI+D program), Universidad Nacional de Lomas de Zamora, the Argentine National Agency for the Promotion of Science and Technology (ANPCyT), and the Argentine National Council for Science and Technology (CONICET).

Association between androgen receptor expression, Ki-67 and the 21-gene recurrence score in non-metastatic, lymph node-negative, estrogen receptor-positive and HER2-negative breast cancer

2015 ◽  
Vol 68 (10) ◽  
pp. 839-843 ◽  
Author(s):  
Francisco E Vera-Badillo ◽  
Martin C Chang ◽  
Gordana Kuruzar ◽  
Alberto Ocana ◽  
Arnoud J Templeton ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Estrogen Receptor ◽  
Lymph Node ◽  
Androgen Receptor ◽  
Statistical Significance ◽  
Multivariable Analysis ◽  
Recurrence Score ◽  
Receptor Expression ◽  
Ki 67 ◽  
Node Negative

BackgroundThe mechanisms underlying the favourable prognosis of androgen receptor (AR) expression in breast cancer are unknown.MethodsThe associations between the 21-gene recurrence score (RS), AR, grade, mitotic score, Ki-67 and estrogen receptor (ER) and progesterone receptor (PgR) expression were explored in sequential women with lymph node-negative, ER-positive and HER2-negative breast cancer. Statistical significance of this exploratory study was defined as p<0.10.ResultsAnalysis comprised 70 women. Most tumours had high AR expression (97% had scores >3). Median RS was 15 (range 1–53). AR expression showed a minimally significant positive correlation with ER (R=0.37), but no correlation with Ki-67 (R=−0.18). In univariable analysis, AR (p=0.01), ER (p<0.001) and PgR (p<0.001) had significant negative associations with RS. Ki-67 (p=0.16), grade (p=0.40) and mitotic score (p=0.23) showed no association with RS. Multivariable analysis showed similar associations.ConclusionsAR is associated with lower RS, but not with Ki-67.

scholarly journals Androgen Receptor Expression and Bicalutamide Antagonize Androgen Receptor Inhibit β-Catenin Transcription Complex in Estrogen Receptor-Negative Breast Cancer

2017 ◽  
Vol 43 (6) ◽  
pp. 2212-2225 ◽  
Author(s):  
Rui Huang ◽  
Jiguang Han ◽  
Xiaoshuan Liang ◽  
Shanshan Sun ◽  
Yongdong Jiang ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Estrogen Receptor ◽  
Androgen Receptor ◽  
Receptor Expression ◽  
Breast Cancer Cell Lines ◽  
Ki 67 ◽  
Cancer Subtypes ◽  
Invasive Ductal Cancer ◽  
Mechanistic Basis

Background/Aims: Little is known about the potential mechanism of action for androgen receptor (AR) targeting treatment in estrogen receptor (ER)-negative breast cancer. This study aimed to evaluate AR status and its prognosis in four breast cancer subtypes. Bicalutamide has been identified as an AR antagonist and used for treating AR+/ER- breast cancer in a phase II trial. Our studies will clarify its mechanism in breast cancer treatment. Methods: A total of 510 consecutive cases of invasive ductal cancer (IDC) were evaluated in this study. The expression of AR was analyzed by immunohistochemistry and compared with patient survival, and its implications were evaluated in four subtypes of IDC. We examined bicalutamide as an AR antagonist to inhibit proliferation and increased apoptosis in AR+/ER- breast cancer cell lines. We explored the tumor suppressive functions of bicalutamide in vitro and vivo and its related mechanisms in AR+/ER- breast cancer. Results: AR expression was related to that of ER (P<0.001), PR (P<0.001), Her2 (P=0.017), Ki-67(P=0.020) and to four subtypes (P<0.001). AR retained independent prognostic signifcance (P=0.007, ER- cases; P=0.001, ER+ cases; P=0.001, total cases). We found that bicalutamide significantly decreased viability and increased apoptosis in vitro and vivo. The mechanistic analysis revealed that bicalutamide blocked androgen-stimulated oncogenic AR and Wnt/β-catenin signaling and inhibited the growth of AR+/ER- breast cancer. Conclusion: Our studies provide novel insights into bicalutamide as an antagonist of AR function in AR+/ER- breast cancer and reveal the mechanistic basis for targeting AR as a therapeutic opportunity for patients with AR+/ER- breast cancer.

Expression of Bcl-2 and 3-ß hydroxysteroid dehydrogenase protein during oocyte and follicle development in fetal and post-natal pig ovaries

1999 ◽  
Vol 11 (8) ◽  
pp. 463 ◽  
Author(s):  
Wesley M. Garrett ◽  
H. David Guthrie
Keyword(s):  
Cell Proliferation ◽  
Germ Cell ◽  
Germ Cells ◽  
Granulosa Cells ◽  
Stromal Cells ◽  
Post Partum ◽  
Nuclear Antigen ◽  
Ki 67 ◽  
Primordial Follicles ◽  
Small Clusters

The fetal and post-natal development of the pig ovary involves both proliferation and apoptotic loss of germ cells, follicle formation and growth, and the initiation of oocyte meiotic maturation. The present study measured the expression of the proto-oncogene Bcl-2 immunohistochemically on paraffin sections of pig ovaries to determine its relationship with folliculogenesis on Days 50 and 80 post coitum (p.c.) and on Days 1, 21, and 56 post partum (p.p.). The expression of the steroidogenic enzyme 3β-hydroxy-steroid dehydrogenase (3βHSD) was used to determine the lineages of the cells forming the ovarian follicles, and the expression of the cell proliferation-associated nuclear antigen Ki-67 was used to determine germ cell proliferation and the initiation of follicle growth. Expression of Ki-67 showed that many oogonia were proliferating on Days 50 and 80 p.c. Granulosa cells were more proliferative on Day 56 p.p. than at any other stage; Ki-67 was expressed in 70% of growing follicles and granulosa cells had a 3% mean staining index per section. Less than 4% of germ cells and follicles had morphological signs of degeneration during the period of the study. Bcl-2 was most abundant on Days 21 p.p. and 56 p.p.; staining was localized to stromal cells among follicles and in small clusters in the cortical–medullary junction (CMJ). 3βHSD staining on Day 50 p.c. was seen in cords of stromal cells within the medulla of the ovary, and in the stromal cells investing the oogonial nests. On Days 80 p.c., 1 p.p., 21 p.p., and 56 p.p., 3βHSD was expressed in the granulosa cells of primary or primordial follicles at the CMJ. Production of Bcl-2 by somatic cells may support germ cell and preantral follicle survival.

scholarly journals Developmental Changes of the Ovary in Neonatal Cotton Rat (Sigmodon hispidus)

2021 ◽  
Vol 11 ◽  
Author(s):  
Md. Rashedul Islam ◽  
Osamu Ichii ◽  
Teppei Nakamura ◽  
Takao Irie ◽  
Md. Abdul Masum ◽  
...  
Keyword(s):  
Ovarian Development ◽  
Developmental Changes ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Sigmodon Hispidus ◽  
Ki 67 ◽  
Primordial Follicles ◽  
Forkhead Box ◽  
Transmission Electron ◽  
Cotton Rats ◽  
Almost All

The reproductive characteristics and ovarian development in cotton rats (Sigmodon hispidus, CRs) are unclear, although CRs are commonly used as animal models in biomedical research. We previously reported that young (6–8 weeks) CRs showed multi-oocyte follicles (MOFs) and double nucleated oocytes (DNOs) in different stages of follicles. The developmental changes in neonatal CR ovaries were investigated in the present study and were compared with our findings in previous studies of unique phenotypes, particularly in oocytes. CR ovaries at postnatal days (PND) 0, 4, and 7 were obtained from the Hokkaido Institute of Public Health. Samples were analyzed by light and transmission electron microscopy. The general histology and folliculogenesis in CR ovaries were similar to those in other experimental rodents. However, DNOs were observed in all age categories and were frequently observed in primordial follicles, whereas MOFs started to develop from PND4 with greater frequency in primary follicles. Almost all developing follicles expressed DEAD (Asp-Glu-Ala-Asp) box polypeptide 4 and forkhead box L2, which are representative markers of oocytes and follicular epithelial cells, respectively. Ki-67 staining demonstrated the proliferative activity of granulosa cells, but not of oocytes, in follicles. Moreover, rapid folliculogenesis of CR due to a small number of apoptotic oocytes was suggested, based on results of the terminal deoxynucleotidyl transferase dUTP nick end labeling assay, confirming the formation of DNOs or MOFs. These findings clarify the development of unique phenotypes of neonatal CR ovaries and support it as a useful model to better understand folliculogenesis and oocytogenesis as well as their abnormalities in humans and other animals.

Immunohistochemical study of Ki-67 protein, androgen receptor, and estrogen receptor beta in testicular tissues of male pigs immunocastrated with different times of GnRH vaccination

2018 ◽  
Vol 47 (5) ◽  
pp. 475-480 ◽  
Author(s):  
Kongkiat Srisuwatanasagul ◽  
Nawarus Prapaiwan ◽  
Sayamon Srisuwatanasagul ◽  
Annop Kunavongkrit ◽  
Atthaporn Roongsitthichai
Keyword(s):  
Estrogen Receptor ◽  
Androgen Receptor ◽  
Immunohistochemical Study ◽  
Estrogen Receptor Beta ◽  
Ki 67 ◽  
Receptor Beta

Changes of tumor infiltrating lymphocyte subtypes before and after neoadjuvant endocrine therapy in estrogen receptor-positive breast cancer patients – an immunohistochemical study of cd8+ and foxp3+ using double immunostaining with correlation to the pathobiological response of the patients

2012 ◽  
Vol 27 (4) ◽  
pp. 295-304 ◽  
Author(s):  
Monica Sm Chan ◽  
Lin Wang ◽  
Saulo Ja Felizola ◽  
Takayuki Ueno ◽  
Masakazu Toi ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Estrogen Receptor ◽  
Endocrine Therapy ◽  
Stromal Cells ◽  
Breast Cancer Patients ◽  
Neoadjuvant Endocrine Therapy ◽  
Ki 67 ◽  
Before And After ◽  
Estrogen Receptor Positive ◽  
Positive Breast Cancer

Tumor-stromal interactions involve continuous crosstalk and interactions among different cell types and play pivotal roles in tumorigenesis, tumor development, disease progression, subsequent metastasis, and also tumor response to therapeutic agents. Tumor infiltrating lymphocytes (TILs) are important components of these tumor-stromal interactions. Specific TIL subtypes are known to be involved in the clinical course of individual patients. However, the status of TILs following endocrine therapy has not been studied in breast cancer patients. We evaluated the alterations of TIL subtypes in a cohort of East Asian patients with estrogen receptor-positive breast cancer during the course of neoadjuvant steroidal aromatase inhibitor (AI) therapy, using double immunohistochemical staining of CD8+ and T regulatory cells (Treg) or Foxp3+, yielding the CD8+/Treg ratio in individual patients. Changes in CD8+/Treg ratio before and after therapy were then correlated with pathobiological responses of individual patients based upon alterations of the Ki-67 labeling index (LI). A significant increase in the CD8+/Treg ratio was detected in responders (p=0.028) but not in non-responders, which may reflect the dynamic process in which the host immune response to tumor antigens changed in consequence of an interaction between tumor and stromal cells in its microenvironment following estrogen depletion caused by the AI. The CD8+/Treg ratio in breast cancer tissue can be a potential surrogate marker in surgical pathology specimens for predicting responses to neoadjuvant endocrine therapy, not only incorporating features of carcinoma cells as in Ki-67 LI but also those of adjacent stromal cells in the tumor microenvironment, especially in the early stage of treatment prior to any detectable clinical and/or histopathological changes.

scholarly journals Gene expression and protein localisation for activin-A, follistatin and activin receptors in goat ovaries

2004 ◽  
Vol 183 (2) ◽  
pp. 405-415 ◽  
Author(s):  
J R V Silva ◽  
R van den Hurk ◽  
H T A van Tol ◽  
B A J Roelen ◽  
J R Figueiredo
Keyword(s):  
Mrna Expression ◽  
Granulosa Cells ◽  
Activin A ◽  
Surface Epithelium ◽  
Corpora Lutea ◽  
Rt Pcr ◽  
Primordial Follicles ◽  
Antral Follicles ◽  
Theca Cells ◽  
Activin Receptors

We studied the protein and mRNA expression of activin-A, follistatin and activin receptors in goat ovaries to find evidence of their possible role in ovarian activity, particularly in the various stages of follicle development. Ovaries of cyclic goats were collected and then either fixed in paraformaldehyde for immunohistochemical localisation of activin-A, follistatin, activin receptors IIA/B (ActR-IIA/B) and IA (ActR-IA) proteins or used to obtain samples to demonstrate mRNA expression of activin-A (βA subunit), follistatin, ActR-IIA, -IIB, -IA and -IB, using RT-PCR. For this latter goal, primordial, primary and secondary follicles were isolated mechanically, washed to remove the stromal cells and then used for RT-PCR. In addition, oocytes, cumulus, mural granulosa and theca cells from small (<3 mm) and large (3–6 mm) antral follicles, luteal cells and surface epithelium were collected to study mRNA expression. Activin-A and follistatin proteins were found in oocytes of all follicle classes, granulosa cells from the primary follicle stage onwards, theca cells of antral follicles, corpora lutea and ovarian surface epithelium. In antral follicles, these proteins were detected both in cumulus and mural granulosa cells. ActR-IIA/B protein was found at the same follicular sites, and also in granulosa cells of primordial follicles onward. The localisation of ActR-IA corresponded with that of ActR-IIA/B, but the former protein was absent in the theca of large antral follicles. The mRNAs for activin-A (βA subunit), follistatin, and ActR-IIA, -IIB, -IA and -IB were detected at all follicular and cellular types studied, except that ActR-IIB was not found in follicles that had not developed an antrum yet. In conclusion, in goat ovaries, transcripts of activin-A (βA subunit), its receptors and its binding protein follistatin are expressed and their proteins formed at all follicular stages and in corpora lutea. These findings indicate a role of activin-A in the local regulatory system during the entire follicular development and during luteal activity.

scholarly journals Expression of Progesterone Receptors A and B in the Mouse Ovary during the Estrous Cycle

Endocrinology ◽  
2004 ◽  
Vol 145 (7) ◽  
pp. 3487-3494 ◽  
Author(s):  
Natalie Gava ◽  
Christine L. Clarke ◽  
Karen Byth ◽  
Rebecca L. Arnett-Mansfield ◽  
Anna deFazio
Keyword(s):  
Monoclonal Antibodies ◽  
Stromal Cells ◽  
Ovarian Function ◽  
Progesterone Receptors ◽  
Cellular Distribution ◽  
Mouse Ovary ◽  
Functional Activities ◽  
Antral Follicles ◽  
Theca Cells ◽  
Preovulatory Follicles

Abstract Progesterone plays a central role in the regulation of ovarian function. The progesterone receptor (PR) has been shown to be essential for ovulation because mice lacking PR fail to ovulate and are infertile. PR is expressed as two isoforms, PRA and PRB, which have been shown to have different functional activities. In this study, we investigated the cellular distribution of PRA and PRB in the ovaries and oviducts of cycling mice using immunohistochemistry with isoform-specific monoclonal antibodies. In the ovary, on the evening of proestrus before ovulation, both the granulosa and theca cells of the preovulatory follicles expressed both PR isoforms. PRA and PRB staining was also observed in the theca cells of preantral and antral follicles, whereas only PRB was observed in the granulosa cells of primary, preantral, and antral follicles and in the corpus luteum. In the oviduct, PRA was the predominant isoform observed, expressed in both the epithelial and stromal cells, whereas PRB was only detected in the epithelial cells. The differences in PRA and PRB localization in the ovary and oviduct may reflect diverse functions for PRA and PRB in reproductive tissues and may have important implications in understanding the mechanisms of progesterone action.

Changes in the Proliferation/Apoptosis Balance in the Bovine Ovary: A Key Early Event in Follicular Persistence

2017 ◽  
Vol 204 (5-6) ◽  
pp. 314-325 ◽  
Author(s):  
E. Matías Belotti ◽  
Antonela F. Stassi ◽  
Melisa M.L. Velázquez ◽  
Pablo U. Díaz ◽  
Belkis E. Marelli ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Granulosa Cells ◽  
Caspase 3 ◽  
Control Group ◽  
Early Event ◽  
Ki 67 ◽  
Antral Follicles ◽  
Theca Cells ◽  
Bovine Ovary ◽  
Bcl2 Expression

The objective of this work was to evaluate proliferation and apoptosis in the bovine ovary in a model of follicular persistence induced by low levels of progesterone to detect incipient changes during cystic ovarian disease development on the expected day of ovulation (day 0) and after 5, 10, and 15 days of follicular persistence. We analyzed cell proliferation by evaluating the expression of Ki-67 and apoptosis by evaluating caspase-3, BAX, and BCL2 expression. Proliferation was similar in the granulosa and theca cells of antral follicles in the P0 group (treated with progesterone up to the expected day of ovulation) and in the control group. A decrease in cell proliferation was detected after 5 days of persistence (P5) in relation to P0 (p < 0.05). Similar changes were found in the granulosa cells of the persistent follicles in relation to the control group (p < 0.05). Caspase-3 expression was similar in granulosa cells of antral follicles at early stages of persistence, with an increase after 15 days of persistence (p < 0.05). In the granulosa cells of group P10 (10 days of persistence), caspase-3 expression was reduced relative to that of antral follicles from the control group (p < 0.05). BCL2 expression was higher in granulosa cells of the persistent follicles of group P0 relative to the control follicles, with no changes in BAX expression, which was increased in persistent follicles of group P15 (p < 0.05). Similar results were observed in theca cells at initial stages of persistence. The results show that, initially, proliferation is maintained with low apoptosis and an increase in cell survival.

Expression of angiotensin II receptors in the caprine ovary and improvement of follicular viability in vitro

Zygote ◽  
2015 ◽  
Vol 24 (4) ◽  
pp. 568-577 ◽  
Author(s):  
J.B. Bruno ◽  
I.B. Lima-Verde ◽  
J.J.H. Celestino ◽  
L.F. Lima ◽  
M.H.T. Matos ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Ultrastructural Analysis ◽  
Mrna Levels ◽  
Ang Ii ◽  
Primordial Follicles ◽  
Preantral Follicles ◽  
Antral Follicles ◽  
Theca Cells ◽  
Cumulus Oocyte Complex

SummaryThis study aimed to evaluate mRNA levels of angiotensin II (ANG II) receptors (AGTR1 and AGTR2) in caprine follicles and to investigate the influence of ANG II on the viability and in vitro growth of preantral follicles. Real-time polymerase chain reaction (PCR) was used to quantify AGTR1 and AGTR2 mRNA levels in the different follicular stages. For culture, caprine ovaries were collected, cut into 13 fragments and then either directly fixed for histological and ultrastructural analysis (fresh control) or placed in culture for 1 or 7 days in α-minumum essential medium plus (α-MEM+) with 0, 1, 5, 10, 50 or 100 ng/ml ANG II. Then, the fragments were destined to morphological, viability and ultrastructural analysis. The results showed that primordial follicles had higher levels of AGTR1 and AGTR2 mRNA than secondary follicles. Granulosa/theca cells from antral follicles had higher levels of AGTR1 mRNA than their respective cumulus–oocyte complex (COCs). After 7 days of culture, ANG II (10 or 50 ng/ml) maintained the percentages of normal follicles compared with α-MEM+. Fluorescence and ultrastructural microscopy confirmed follicular integrity in ANG II (10 ng/ml). In conclusion, a high expression of AGTR1 and AGTR2 is observed in primordial follicles. Granulosa/theca cells from antral follicles had higher levels of AGTR1 mRNA. Finally, 10 ng/ml ANG II maintained the viability of caprine preantral follicles after in vitro culture.

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