125 Transcriptomic changes in bovine ovarian cortex in response to FSH signaling

2020 ◽  
Vol 32 (2) ◽  
pp. 189
Author(s):  
J. Candelaria ◽  
B. Rabaglino ◽  
A. Denicol
Keyword(s):  
Time Course ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Signalling Pathways ◽  
Biological Functions ◽  
Cortical Cells ◽  
Preantral Follicles ◽  
Antral Follicles ◽  
Ovarian Cortex ◽  
Over Time

Preantral follicles serve as a reservoir of female gametes that could be used in assisted reproductive technologies in humans, livestock, and endangered animals. Invitro culture of ovarian cortex is a widely used method to grow preantral follicles. Follicle-stimulating hormone (FSH) is often added to the culture medium as a folliculogenesis-promoting factor. The roles of FSH in antral follicles is well known; however, the effects of FSH in preantral follicles and indirectly in the ovarian cortical cells is largely unknown. The aim of this study was to determine the transcriptomic responses of the ovarian cortex containing preantral follicles to FSH signalling over time. In 3 biological replicates, small strips of bovine ovarian cortex (10×5mm) were dissected from the medulla and evaluated under a stereomicroscope for removal of all visible antral follicles. Resulting cortical strips were cultured in defined medium with human-recombinant FSH or vehicle for 2 or 4 days at 38.5°C and 5% CO2. The RNA was isolated and subjected to cDNA library preparation and 3′-Tag RNA sequencing. Sequencing data analysis was performed using the edgeR and maSigPro packages (Bioconductor-R). Using a time-course analysis, genes up- or downregulated 2-fold or more and associated with an FDR<0.05 were considered differentially expressed (DEG) and were further analysed with NetworkAnalyst software. We found 252 DEG over time in response to FSH. In FSH-treated samples, significantly enriched biological functions from upregulated genes were associated with glycolysis, gluconeogenesis, carbon metabolism, and biosynthesis of amino acids. In contrast, significantly enriched biological functions from downregulated genes found in FSH-treated samples included phagosome and necroptosis. The germ cell markers BMP15, DAZL, DDX4, GDF9, and ZP2/ZP3 were expressed but unchanged by FSH, suggesting the presence of similar numbers of oocytes between samples. The gene B4GALT2, previously reported as a granulosa cell marker, was upregulated in FSH-treated samples at Day 4. The follicular marker RAB23 was expressed in all samples and not changed by FSH. One interesting finding was upregulation of MAPK signalling (represented by the genes MAPKAPK3, ELK4, MKNK2, and TGFB3) in response to FSH signalling, with no change in expression of the cAMP-response element responsive genes CYP19A1 and INHA. Together, these data indicate that FSH stimulates energy metabolism in ovarian cortical cells and represses negative cell function activity. We conclude that these responses are mostly mediated by granulosa cells, because the FSH receptor is not appreciably expressed in the ovarian cortex stroma. Moreover, the data suggest that FSH may utilise alternative signalling pathways, such as MAPK, in early follicles. This information enhances our understanding of FSH signalling pathways in the ovarian cortex, mediated by preantral follicles to create a positive environment for folliculogenesis.

Characterization of isolated bovine preantral follicles based on morphology, diameter and cell number

Zygote ◽  
2020 ◽  
Vol 28 (2) ◽  
pp. 154-159
Author(s):  
Juliana I. Candelaria ◽  
Anna C. Denicol
Keyword(s):  
Granulosa Cells ◽  
Developmental Stages ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Cell Number ◽  
Culture Conditions ◽  
Preantral Follicles ◽  
Secondary Stage

SummaryPreantral follicles are a potential reservoir of oocytes to be used in assisted reproductive technologies. With the increasing interest in developing techniques to grow preantral follicles in vitro, and as the bovine emerges as an appropriate model species to understand human folliculogenesis, the establishment of an accurate classification of developmental stages is needed. Classification of bovine preantral follicles has been mostly based on histological analysis and estimation models, which may not translate well to correctly characterize preantral follicles isolated from the ovary. In this study, we classified bovine preantral follicles by morphology upon isolation, determined diameter and number of granulosa cells by direct counting, and compared our results with previous studies reporting bovine preantral follicle classification. Follicles were isolated via homogenization of ovary tissue and classified into primary, early secondary and secondary stage based on morphology and number of layers of granulosa cells. Diameter was individually measured and Hoechst 33342 was used as a nuclear stain to count granulosa cells. We found that follicles classified by morphology into primary, early secondary, and secondary had different mean diameter and cell number (P < 0.01); cell number and diameter were positively correlated, as were cell density and cell number in each developmental stage (P < 0.01). Results obtained here were mostly in agreement with previous classifications based on histological sections and on isolated follicles, with some discrepancies. The present data add accuracy to classification of bovine preantral follicles that is critical to optimize culture conditions to produce developmentally competent oocytes.

scholarly journals “Man Cannot Exist Alone”: The Challenge of Health Care to Honor the Role of Women in Society

2018 ◽  
Vol 85 (4) ◽  
pp. 327-330
Author(s):  
Jonathan Scrafford
Keyword(s):  
Health Care ◽  
Reproductive Health ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Natural Family Planning ◽  
Social And Emotional ◽  
Emotional Bonds ◽  
Parents And Children ◽  
Women's Role ◽  
Over Time

Women’s roles in society are changing. While most of those changes recognize and enhance the contributions of feminine ingenuity to human development, some threaten to isolate women physically, socially, and emotionally. Developments in reproductive health care, and the writings of Pope Saint John Paul II, offer lenses by which to evaluate the shifting landscape of women’s role in society. On the one hand, practices such as contraception, abortion, surrogacy, and assisted reproductive technologies over time will weaken the physical, social, and emotional bonds that procreation has held between man and woman, parents and children, and families and society. On the other hand, the expansion of different modes of natural family planning and pregnancy support centers offers to preserve those bonds. Summary: Women’s role in families, and therefore society, is invaluable. Several approaches to reproductive health offered by medicine may isolate women over time, and some evidences suggest we are already seeing that effect. Other approaches to women’s health may be able to preserve the physical, emotional, and social bonds that integrate women to the family, and therefore society.

Impacts of and interactions between environmental stress and epigenetic programming during early embryo development

2015 ◽  
Vol 27 (8) ◽  
pp. 1125 ◽  
Author(s):  
Michael J. Bertoldo ◽  
Yann Locatelli ◽  
Christopher O'Neill ◽  
Pascal Mermillod
Keyword(s):  
Embryo Development ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Culture Conditions ◽  
Early Embryo ◽  
Signalling Pathways ◽  
Epigenetic Programming ◽  
Altered Gene ◽  
Vitro Development

The processes of assisted reproductive technologies (ART) involve a variety of interventions that impact on the oocyte and embryo. Critically, these interventions cause considerable stress and coincide with important imprinting events throughout gametogenesis, fertilisation and early embryonic development. It is now accepted that the IVM and in vitro development of gametes and embryos can perturb the natural course of development to varying degrees of severity. Altered gene expression and, more recently, imprinting disorders relating to ART have become a focused area of research. Although various hypotheses have been put forward, most research has been observational, with little attempt to discover the mechanisms and periods of sensitivity during embryo development that are influenced by the culture conditions following fertilisation. The embryo possesses innate survival factor signalling pathways, yet when an embryo is placed in culture, this signalling in response to in vitro stress becomes critically important in mitigating the effects of stresses caused by the in vitro environment. It is apparent that not all embryos possess this ability to adequately adapt to the stresses experienced in vitro, most probably due to an inadequate oocyte. It is speculated that it is important that embryos use their survival signalling mechanisms to maintain normal epigenetic programming. The seeming redundancy in the function of various survival signalling pathways would support this notion. Any invasion into the natural, highly orchestrated and dynamic process of sexual reproduction could perturb the normal progression of epigenetic programming. Therefore the source of gametes and the subsequent culture conditions of gametes and embryos are critically important and require careful attention. It is the aim of this review to highlight avenues of research to elucidate the effects of stress and the relationship with epigenetic programming. The short- and long-term health and viability of human and animal embryos derived in vitro will also be discussed.

scholarly journals Comparative characteristics of patients with infertility when applying melatonin in complex preparation for assisted reproductive technologies

2021 ◽  
Vol 25 (3 (99)) ◽  
pp. 149-156
Author(s):  
V. Yuzko ◽  
O. Yuzko
Keyword(s):  
Ovarian Reserve ◽  
Assisted Reproductive Technologies ◽  
Ultrasound Examination ◽  
Reproductive Technologies ◽  
Control Group ◽  
Antral Follicles ◽  
The Third ◽  
Significant Difference ◽  
Ovulation Stimulation ◽  
Infertility Patients

Оbjective of the study was to conduct a retrospective comparative characterization of patients with infertility who took or did not take melatonin with assisted reproductive technologies (ART).Materials and methods. In our study, we examined 89 women. The first (control) group included 13 healthy women oocyte donors who got pregnant on their own and gave birth to their own healthy children, the second group - 33 patients with infertility, who took 3 mg of the preparation "Vita-melatonin" produced by "Kyiv Vitamin Plant" at the same time before bedtime, two weeks before and during ovulation stimulation, the third group - 43 patients with infertility who did not take melatonin preparation before and during ovulation stimulation. There were no women who worked night shifts among the patients. Medical documentation of women of the control group and those with infertility, data of gynecological, ultrasound examination, hormones blood were analyzed. Ultrasound examination of the pelvic organs was performed on all patients with the device "Mindray DC-80 X-Insight", and measurements were performed using a transvaginal sensor. The thickness and structure of the endometrium were evaluated, and the number of antral follicles (NAF) ranging in size from 2 to 10 mm was counted in each ovary. All patients were tested for serum levels of the anti-mullerian hormone (AMG), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), prolactin (PRL), progesterone (P), thyroid-stimulating hormone (TSH), triiodothyronine (T4).Results. The average age of women in the first (control) group was 27.08 ± 12.38 years, the second (taking melatonin) - 33.12 ± 8.18 years, the third (not taking melatonin) - 30.95 ± 7.07 years > 0.05), i. e. the age of the patients of the examined groups was equal. It should be noted that in the studied patients of both groups, the occurrence of primary infertility exceeded secondary infertility 2.7 times in the second group (p < 0.05) and 1.7 times in the third (p < 0.05). Infertility factors such as reduced ovarian reserve, habitual miscarriage and infertility of unknown origin were more common in patients of the second group, and endometriosis, tubal factor and male factor in the third, although the difference was not significant. The available extragenital pathology did not differ in the patients of the examined groups. The number of antral follicles was significantly higher in both ovaries of women in the control group compared with patients of the second and third groups. While the thickness of the endometrium did not differ significantly in groups, although in women of the control group it was slightly less. Regarding the study of hormonal status, it should be noted that we did not find a significant difference in the levels of hormones in the blood of women we examined. Exceptionally, there was a significant difference (p < 0.001) in progesterone content between the second (0.62 ± 0.052 nmol/l) and third (181.63 ± 13.87 nmol/l) groups. Also, the patients of the third group had significantly (p < 0.05) higher levels of FSH in blood (8.25 ± 0.63 mUn/ml) compared with the control group (4.93 ± 0.69 mUn/ml).Conclusions. The examined women in the control group, as well as infertility patients who received melatonin two weeks before the expected menstruation and during ovulation stimulation, and infertility patients who did not receive this preparation in similar programs, did not differ in age, occurrence of primary and secondary infertility, the factor that led to infertility, concomitant extragenital pathology, ovarian reserve and hormone levels of the reproductive panel. That is, they were equal in our study.

scholarly journals Gene expression differences induced by equimolar low doses of LH or hCG in combination with FSH in cultured mouse antral follicles

2012 ◽  
Vol 215 (2) ◽  
pp. 269-280 ◽  
Author(s):  
Ingrid Segers ◽  
Tom Adriaenssens ◽  
Sandra Wathlet ◽  
Johan Smitz
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Gene Selection ◽  
Assisted Reproductive Technologies ◽  
Day Treatment ◽  
Reproductive Technologies ◽  
Human Menopausal Gonadotropin ◽  
Follicle Growth ◽  
Gnrh Analog ◽  
Antral Follicles

In a natural cycle, follicle growth is coordinated by FSH and LH. Follicle growth stimulation in Assisted Reproductive Technologies (ART) requires antral follicles to be exposed to both FSH and LH bioactivity, especially after GNRH analog pretreatment. The main aim was to detect possible differences in gene expression in granulosa cells after exposing the follicle during antral growth to LH or hCG, as LH and hCG are different molecules acting on the same receptor. Effects of five gonadotropin treatments were investigated for 16 genes using a mouse follicle culture model. Early (day 6) antral follicles were exposed to high recombinant FSH combined or not with equimolar concentrations of recombinant LH (rLH) or recombinant hCG (rhCG) and to highly purified human menopausal gonadotropin (HP-hMG) for 6 h, 12 h, or 3 days. Expression differences were tested for genes involved in steroidogenesis:Mvk,Lss,Cyp11a1,Hsd3b1,Cyp19a1,Nr4a1, andTimp1; final granulosa differentiation:Lhcgr,Oxtr,Pgr,Egfr,Hif1a, andVegfa; and cytokines:Cxcl12,Cxcr4, andSdc4.Lhcgrwas present and upregulated by gonadotropins.Nr4a1,Cxcl12, andCxcr4showed a different expression pattern if LH bioactivity was added to high FSH in the first hours after exposure. However, no signs of premature luteinization were present even after a 3-day treatment as shown byCyp19a1,Oxtr,Pgr, andEgfrand by estrogen and progesterone measurements. The downstream signaling by rhCG or rLH through the LHCGR was not different for this gene selection. Granulosa cells from follicles exposed to HP-hMG showed an enhanced expression level for several genes compared with recombinant gonadotropin exposure, possibly pointing to enhanced cellular activity.

Re-imagining Reproduction: Unsettling Metaphors in the History of Imperial Science and Commercial Surrogacy in India

Somatechnics ◽  
2015 ◽  
Vol 5 (1) ◽  
pp. 88-103 ◽  
Author(s):  
Kalindi Vora
Keyword(s):  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
The Body ◽  
Body Parts ◽  
Colonial History ◽  
Women's Bodies ◽  
Commercial Surrogacy ◽  
Art Practices ◽  
History Of ◽  
Travel Industry

This paper provides an analysis of how cultural notions of the body and kinship conveyed through Western medical technologies and practices in Assisted Reproductive Technologies (ART) bring together India's colonial history and its economic development through outsourcing, globalisation and instrumentalised notions of the reproductive body in transnational commercial surrogacy. Essential to this industry is the concept of the disembodied uterus that has arisen in scientific and medical practice, which allows for the logic of the ‘gestational carrier’ as a functional role in ART practices, and therefore in transnational medical fertility travel to India. Highlighting the instrumentalisation of the uterus as an alienable component of a body and subject – and therefore of women's bodies in surrogacy – helps elucidate some of the material and political stakes that accompany the growth of the fertility travel industry in India, where histories of privilege and difference converge. I conclude that the metaphors we use to structure our understanding of bodies and body parts impact how we imagine appropriate roles for people and their bodies in ways that are still deeply entangled with imperial histories of science, and these histories shape the contemporary disparities found in access to medical and legal protections among participants in transnational surrogacy arrangements.

Ultrastructure of the basal lamina of bovine ovarian follicles and its relationship to the membrana granulosa

Reproduction ◽  
2000 ◽  
pp. 221-228 ◽  
Author(s):  
HF Irving-Rodgers ◽  
RJ Rodgers
Keyword(s):  
Basal Lamina ◽  
Granulosa Cells ◽  
Light And Electron Microscopy ◽  
Single Layer ◽  
Antral Follicle ◽  
Basal Cells ◽  
Preantral Follicles ◽  
Antral Follicles ◽  
Cellular Processes ◽  
Innermost Layer

Different morphological phenotypes of follicular basal lamina and of membrana granulosa have been observed. Ten preantral follicles (< 0. 1 mm), and 17 healthy and six atretic antral follicles (0.5-12 mm in diameter) were processed for light and electron microscopy to investigate the relationship the between follicular basal lamina and membrana granulosa. Within each antral follicle, the shape of the basal cells of the membrana granulosa was uniform, and either rounded or columnar. There were equal proportions of follicles </= 4 mm in diameter with columnar basal cells and with rounded basal cells. Larger follicles had only rounded basal cells. Conventional basal laminae of a single layer adjacent to the basal granulosa cells were observed in healthy follicles at the preantral and antral stages. However, at the preantral stage, the conventional types of basal lamina were enlarged or even partially laminated. A second type of basal lamina, described as 'loopy', occurred in about half the preantral follicles and in half the antral follicles </= 4 mm diameter. 'Loopy' basal laminae were not observed in larger follicles. 'Loopy' basal laminae were composed of basal laminae aligning the basal surface of basal granulosa cells, but with additional layers or loops often branching from the innermost layer. Each loop was usually < 1 microm long and had vesicles (20-30 nm) attached to the inner aspect. Basal cellular processes were also common, and vesicles could be seen budding off from these processes. In antral follicles, conventional basal laminae occurred in follicles with rounded basal granulosa cells. Other follicles with columnar cells, and atretic follicles, had the 'loopy' basal lamina phenotype. Thus, follicles have different basal laminae that relate to the morphology of the membrana granulosa.

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