• Title/Summary/Keyword: Granulosa cells

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Analysis of protein-protein interaction network based on transcriptome profiling of ovine granulosa cells identifies candidate genes in cyclic recruitment of ovarian follicles

  • Talebi, Reza;Ahmadi, Ahmad;Afraz, Fazlollah
    • Journal of Animal Science and Technology
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    • v.60 no.6
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    • pp.11.1-11.7
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    • 2018
  • After pubertal, cohort of small antral follicles enters to gonadotrophin-sensitive development, called recruited follicles. This study was aimed to identify candidate genes in follicular cyclic recruitment via analysis of protein-protein interaction (PPI) network. Differentially expressed genes (DEGs) in ovine granulosa cells of small antral follicles between follicular and luteal phases were accumulated among gene/protein symbols of the Ensembl annotation. Following directed graphs, PTPN6 and FYN have the highest indegree and outdegree, respectively. Since, these hubs being up-regulated in ovine granulosa cells of small antral follicles during the follicular phase, it represents an accumulation of blood immune cells in follicular phase in comparison with luteal phase. By contrast, the up-regulated hubs in the luteal phase including CDK1, INSRR and TOP2A which stimulated DNA replication and proliferation of granulosa cells, they known as candidate genes of the cyclic recruitment.

Relationship between Sloan-Kettering Virus Expression and Granulosa Cells of Atretic Follicles in the Rat Ovary

  • Kim, Hyun;Matsuwaki, Takashi;Yamanouchi, Keitaro;Nishihara, Masugi;Kim, Sung-Woo;Ko, Yeoung-Gyu;Yang, Boh-Suk
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.341-348
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    • 2011
  • Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. The aim of the present study was to locate Ski protein in rat ovaries in order to predict the possible involvement of Ski in follicular development and atresia. First, expression of c-Ski mRNA in the ovaries of adult female rats was confirmed by RT-PCR. Then, ovaries obtained on the day of estrus were subjected to immunohistochemical analysis for Ski and proliferating cell nuclear antigen (PCNA) in combination with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). Ski was expressed in granulosa cells that were positive for TUNEL, but negative for PCNA, regardless of the shape and size of follicles. Expression of Ski in TUNEL-positive granulosa cells, but not in PCNA-positive granulosa cells, was also verified in immature hypophysectomized rats having a single generation of developing and atretic follicles by treatment with equine chorionic gonadotropin (eCG). These results indicate that Ski is profoundly expressed in the granulosa cells of atretic follicles, but not in growing follicles, and suggest that Ski plays a role in apoptosis of granulosa cells during follicular atresia.

EFFECTS OF HYPOPHYSECTOMY ON PROGESTERONE PRODUCTION IN THE FOLLICULAR GRANULOSA CELLS OF THE JAPANESE QUAIL

  • Mori, M.;Kimora, K.;Yamamuro, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.1 no.3
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    • pp.143-147
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    • 1988
  • In order to investigate the mechanism of regulation of progesterone production, quail were hypophysectomized at various times during the ovulation cycle, and granulose cells were isolated from follicles 4 hr after the operation. They were incubated in vitro at $40^{\circ}C$ with or without LH or dibutyryl cyclic AMP, and the amounts of progesterone produced during 3 hr of incubation were measured by radioimmunoassay. Hypophysectomy at 8 hr or 20 hr before the predicted time of ovulation caused a reduced responsiveness of F1 granulosa cells to exogenous LH or dibutyrul cyclic AMP. Although hypophysectomy at 24 hr before ovulation caused a slight reduction of responsiveness of F1 granulosa cells, the reduction of the progesterone production during the incubation without any stimuli was prominent by the sham operation. These results suggest that the presence of pituitary gland influences the ability of the granulose cells to produce progesterone in response to LH or dibutyryl cyclic AMP.

Determination of Apoptosisin Granulosa-Luteal Cells Obtained from Hyperstimulated Human Ovaries (과배란 유도시 인간 난소로부터 얻어진 과립-황체화 세포의 자연세포사 검정)

  • 양현원
    • Development and Reproduction
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    • v.1 no.1
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    • pp.79-89
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    • 1997
  • Recent studies have demonstrated that apoptotic cell death plays an important role in the mechanism underlying follicular atresia and luteolysis. However, the mechanisms responsible for initiating these processes have not been elucidated. In in vitro fertilization (IVF) programs, it is highly possible that continuous and repeated administration of FSH/hMG and GnRH agonists for the usage of ovarian hyperstimulation may induce apoptotic death of granulosa cells leading to atresia in the human ovarian follicles. The present study was performed to investigate whether FSH/hMG and GnRh agonists used for a longer period in controlled ovarian hyperstimulation has any effect on the apoptosis of granulosa-luteal (GL) cells obtained from hyperstimulated ovaries. To examine apoptotic cell death in the GL cells, cells were stained with acridie orange followed by observed in some of GL cells. Similar but distinct staining of apoptotic GL cells was observed when the cells were examined by using in situ TUNEL method. The healthy-looking cells with normal nuclear morphology were not stained, whereas cells with pyknotic nuclei or with apoptotic nuclei were intensively stained. After examining the ultrastructural features of GL cells by TEM, it was confirmed that the majority of cells seemed to have normal nuclei while GL cells undergoing apoptotic cel death were rarely found. The DNA extracted from GL cells showed a typical pattern of fragmentation following DNA electrophoretic analysis. We have confirmed that the apoptosis occurs in granulosa-luteal cells obtained from hyperstimulated ovaries. Technically, in situ apoptosis detection method is simple and reproducible and is well suited to identify the quality of oocytes retrieved from hyperstimulated ovaries.

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Effects of Steroid Hormone in Avian Follicles

  • Caicedo Rivas, R.E.;Nieto, M. Paz-Calderon;Kamiyoshi, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.4
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    • pp.487-499
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    • 2016
  • The aim of the present study was to examine the effects of testosterone (T) and estradiol-$17{\beta}$ ($E_2$) on the production of progesterone ($P_4$) by granulosa cells, and of the $E_2$ on the production of $P_4$ and T by theca internal cells. In the first experiment, granulosa cells isolated from the largest ($F_1$) and third largest ($F_3$) preovulatory follicle were incubated for 4 h in short-term culture system, $P_4$ production by granulosa cells of both $F_1$ and $F_3$ was increased in a dose-dependent manner by ovine luteinizing hormone (oLH), but not T or $E_2$. In the second experiment, $F_1$ and $F_3$ granulosa cells cultured for 48 h in the developed monolayer culture system were recultured for an additional 48 h with increasing doses of various physiological active substances existing in the ovary, including T and $E_2$. Basal $P_4$ production for 48 h during 48 to 96 h of the cultured was about nine fold greater by $F_1$ granulosa cells than by $F_3$ granulosa cells. In substances examined oLH, chicken vasoactive intestinal polypeptide (cVIP) and T, but not $E_2$, stimulated in a dose-dependent manner $P_4$ production in both $F_1$ and $F_3$ granulosa cells. In addition, when the time course of $P_4$ production by $F_1$ granulosa cells in response to oLH, cVIP, T and $E_2$ was examined for 48 h during 48 to 96 h of culture, although $E_2$ had no effect on $P_4$ production by granulosa cells of $F_1$ during the period from 48 to 96 h of culture, $P_4$ production with oLH was found to be increased at 4 h of the culture, with a maximal 9.14 fold level at 6 h. By contrast, $P_4$ production with cVIP and T increased significantly (p<0.05) from 8 and 12 h of the culture, respectively, with maximal 6.50 fold response at 12 h and 6, 48 fold responses at 36 h. Furthermore, when $F_1$ granulosa cells were precultured with $E_2$ for various times before 4 h culture with oLH at 96 h of culture, the increase in $P_4$ production in response to oLH with a dose-related manner was only found at a pretreatment time of more than 12 h. In the third experiment, theca internal cells of $F_1$, $F_2$ and the largest third to fifth preovulatory follicles ($F_{3-5}$) were incubated for 4 h in short-term culture system with increasing doses of $E_2$. The production of $P_4$ and T by theca internal cells were increased with the addition of $E_2$ of $10^{-6}M$. These increases were greater in smaller follicles. These results indicate that, in granulosa cells of the hen, T may have a direct stimulatory action in the long term on $P_4$ production, and on $E_2$ in long-term action which may enhance the sensitivity to LH for $P_4$ production, and thus, in theca internal cells, $E_2$ in short term action may stimulate the production of $P_4$ and T.

EFFECT OF GRANULOSA AND CUMULUS CELLS ON IN VITRO DEVELOPMENT OF THE BOVINE FOLLICULAR OOCYTES

  • Im, K.S.;Kim, H.J.;Chung, K.M.;Kim, H.S.;Park, K.W.;Niwa, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.4
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    • pp.317-320
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    • 1995
  • The effect of co-culture with cumulus cells and granulosa cells during maturation and development on in vitro developmental potency of follicular oocytes was examined. TCM-199 supplemented with 15% FCS and hormones was used as maturation medium. Sperm from frozen semen was capacitated in modified mTALP medium containing 0.3% BSA, $10{\mu}g/ml$ heparin and 5 mM/ml caffeine. The fertilized embryos were co-cultured on monolayer of cumulus cells or granulosa cells in TCM-199. The embryo co-cultured with cumulus cells showed higher percentage of embryo developed to morula and blastocyst (73.3%) than the embryo co-cultured without cumulus cells (30.8%). The percentage of oocytes developed to morula and blastocyst among cleaved oocytes was significantly (p < 0.05) higher in the oocytes co-cultured with cumulus cells during development (62.4%) than in the oocytes co-cultured with granulosa cells during maturation and with cumulus cells during development (52.3%), and in the oocytes co-cultured with granulose cells during development (52.8%). The results of this study indicate that co-culture of in vitro fertilized embryos with cumulus cells in the development medium increased the rate of embryos developed to morula and blastocyst among cleaved oocytes.

A Study on the Fine Structural Changes of Porcine Ovarian Follicles during Atresia (돼지 난소내 여포의 폐쇄에 따른 미세구조의 변화에 관한 연구)

  • 김문규;이양한김종흡윤용달
    • The Korean Journal of Zoology
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    • v.30 no.4
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    • pp.351-370
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    • 1987
  • This experiment has been done in order to study the correlation between the ultrastructure changes and the atresia phenomenon of the follicles in porcine ovary. The ovaries were assorted according to the presence or absence of corpus luteum. Thereafter, the follicles were classified into normal, pyknotic, necrotic and cystic groups by atretic characteristics on the histological observation, and then their ultrastructures were examined with an electron microscope. The results were as followings. 1. In normal group, granulosa cells represented the ultrastructural characteristics of protein-synthesizing cells. Since the initiation of atresia, the line structure of granulosa cells showed many of the characteristic features of steroid-secreting cells, followed by gradual pyknosis. 2. In necrotic group of the ovary without corpus luteum, the theca interns became hypertrophic and displayed the ultrastructural features of active steroidsecreting cells. But this phenomenon was not seen in the follicles of the ovary with corpus luteum. 3. Degenerative changes of cumulus cells were similar to those of granulosa cells, and the degenerating oocytes showed the degeneration of cellular organelles, cytoplasmic vacuolization and disappearance of microvilli on the surface. The degeneration of granulosa cells tended to procede that of oocytecumulus complex in the follicles of the ovary having no corpus luteum, but this tendency was reversed in the case of presence of corpus luteum. In conclusion, it may be unable to identi(y the initiation of follicular atresia

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Effect of Leptin on the Steroidogenesis of Cultured Human Granulosa Cells (인간 난소의 과립 세포 배양 중 Leptin이 스테로이드 생성에 미치는 영향)

  • Kim, Sei-Kwang;Kim, Myong-Shin;Hwang, Kyung-Joo;Kwon, Hyuck-Chan;Cho, Dong-Jae
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.15-22
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    • 2000
  • Objective: To elucidate the location of leptin and receptors of ovary specimens obtained from patients undergoing hysterectomy by immunohistochemical staining and to determine the effect of leptin on the steroidogenesis of cultured granulosa cells. Method: In the culturing process of the granulosa cells, FSH (1 IU/ml)and leptin (50 ng/ml), IGF-I (50 ng/ml) was administered to each study group (Group I: FSH; Group II: FSH, leptin; Group III: FSH, IGF-I; Group IV: FSH, IGF-I, leptin), and the levels of estradiol, progesterone, androstenedione in the culture media was measured by radioimmunoassay. Statistical analysis was conducted by one-way ANOVA with Scheffe test. Results: The results showed that leptin and leptin receptors were both found to be strongly stained in granulosa and theca cells, and also in some interstitial cells. Leptin receptors were also observed in cultured granulosa cells. While there was no statistically significant difference in the androstnedione concentrations between the groups, estradiol concentrations was significantly decreased in Group IV ($2202.0{\pm}151.14$ pg/ml) compared to Group III ($2859.0{\pm}122.6$ pg/ml), and progesterone concentrations were also significantly decreased in Group II($4696.3{\pm}190.6$ ng/ml) and Group IV ($4517{\pm}206.78$ ng/ml) compared to Group III($5546.0{\pm}179.5$ ng/ml). Conclustion: The study result of this study suggest that leptin is directly involved in the regulation of ovarian functions, in particular steroidogenesis.

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Effects of Bisphenol S on Viability and Reactive Oxygen Species of the Sperm and Ovarian Granulosa Cells in Pigs (Bisphenol S가 돼지정자와 난소내 과립막세포의 생존성과 활성산소에 미치는 영향)

  • Lee, Yu-Sub;Lee, Seunghyung;Yang, Boo-Keun
    • ANNALS OF ANIMAL RESOURCE SCIENCES
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    • v.29 no.4
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    • pp.166-171
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    • 2018
  • The effect of bisphenol S (BPS) on the viability and production of reactive oxygen species (ROS) was studied in boar sperm and ovarian granulosa cells. Boar semen was incubated in Beltsville thawing solution with either 0 or $5{\mu}M$ BPS for 3 and 6 h. The viability of sperm was analyzed by SYBR14/PI doubling staining, and production of ROS was detected. Ovarian granulosa cells were also treated with BPS for 24, 48, and 72 h. Then, cell viability (0, 5, 10, and $20{\mu}M$) and ROS production (only 0 and $5{\mu}M$ BPS) were assessed. The results showed that, BPS decreased sperm viability at 3 and 6 h, and that BPS increased ROS production (p<0.05). Also, BPS reduced the viability of ovarian granulosa cells (p<0.05), and stimulated ROS production (p<0.05). These results suggest that BPS damages sperm activation and ovarian granulosa cells in the reproductive system.

Integrated transcriptomic analysis on small yellow follicles reveals that sosondowah ankyrin repeat domain family member A inhibits chicken follicle selection

  • Zhong, Conghao;Liu, Zemin;Qiao, Xibo;Kang, Li;Sun, Yi;Jiang, Yunliang
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1290-1302
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    • 2021
  • Objective: Follicle selection is an important process in chicken egg laying. Among several small yellow (SY) follicles, the one exhibiting the highest expression of follicle stimulation hormone receptor (FSHR) will be selected to become a hierarchal follicle. The role of lncRNA, miRNA and other non-coding RNA in chicken follicle selection is unclear. Methods: In this study, the whole transcriptome sequencing of SY follicles with different expression levels of FSHR in Jining Bairi hens was performed, and the expression of 30 randomly selected mRNAs, lncRNAs and miRNAs was validated by quantitative real-time polymerase chain reaction. Preliminary studies and bioinformatics analysis were performed on the selected mRNA, lncRNA, miRNA and their target genes. The effect of identified gene was examined in the granulosa cells of chicken follicles. Results: Integrated transcriptomic analysis on chicken SY follicles differing in FSHR expression revealed 467 differentially expressed mRNA genes, 134 differentially expressed lncRNA genes and 34 differentially expressed miRNA genes, and sosondowah ankyrin repeat domain family member A (SOWAHA) was the common target gene of three miRNAs and one lncRNA. SOWAHA was mainly expressed in small white (SW) and SY follicles and was affected by follicle stimulation hormone (FSH) treatment in the granulosa cells. Knockdown of SOWAHA inhibited the expression of Wnt family member 4 (Wnt4) and steroidogenic acute regulatory protein (StAR) in the granulosa cells of prehierarchal follicles, while stimulated Wnt4 in hierarchal follicles. Overexpression of SOWAHA increased the expression of Wnt4 in the granulosa cells of prehierarchal follicles, decreased that of StAR and cytochrome P450 family 11 subfamily A member 1 in the granulosa cells of hierarchal follicles and inhibited the proliferation of granulosa cells. Conclusion: Integrated analysis of chicken SY follicle transcriptomes identified SOWAHA as a network gene that is affected by FSH in granulosa cells of ovarian follicles. SOWAHA affected the expression of genes involved in chicken follicle selection and inhibited the proliferation of granulosa cells, suggesting an inhibitory role in chicken follicle selection.