• Asian-Australasian Journal of Animal Sciences
• /
• v.11 no.5
• /
• pp.486-490
• /
• 1998

The present study examined the follicular populations prior to and during superovulation and investigated their relationship with superovulatory response in crossbred cattle. Eleven animals were administered i.m. 8 doses of Folltropin of 2.5 ml each (1.75 mg/ml) spread over 4 days beginning on Day 10 of oestrous cycle, and 30 and 20 mg Lutalyse was given alongwith the 5th and 6th injections of Folltropin, respectively, to induce luteolysis. The animals were artificially inseminated 48, 60 and 72 h after the first Lutalyse injection. The number of corpora lutea (CL) was recorded by palpation per rectum and embryos were recovered non-surgically on Day 6 (Day 0 day of superoestrus). The ovarian follicular population was examined by transrectal Ultrasonography 15 h prior to and 52 h after the first FSH injection, and then on the day of superoestrus and the day of flushing. The follicles were classfied on the basis of diameter as small (3-5 mm), medium (6-9 mm) and large (${\geq}10mm$). The total number of follicles increased significantly (p < 0.01) from $2.45{\pm}0.35$, 15 h prior to the first FSH injection to $8.09{\pm}1.12$, 52 h after the first FSH injection and then further to $13.27{\pm}1.89 $ on the day of superoestrus. A positive correlation was observed between the number of small follicles 15 h prior to the first FSH injection (r = 0.60, p < 0.05), the number of large follicles 52 h after the first FSH injection (r=0.59, p < 0.05) and the number of CL. The follicular population prior to and during superovulation was, however, not significantly different between high (> 6 CL) and low responders (${\leq}6CL$). The present study suggests that the follicular populations undergo dynamic changes during superovulation and that follicular populations prior to superovulation have a limited application as an indicator of the superovulatory response.

• Animal cells and systems
• /
• v.3 no.4
• /
• pp.385-391
• /
• 1999

Changes in the levels of prostaglandian F$_{2a}$ (PGF$_{2a}$) and E$_2$ (PGE$_2$) in culture medium during in vitro ovulation of Rana dybowskii follicles were examined. The ovulation was induced by frog pituitary homogenate (FPH) or TPA (12-O-tetradecanoylphorbol-13-acetate, a protein kinase activator) and the levels of PGs were measured by radioimmunoassay. When the ovarian follicles were cultured, only a few oocytes were ovulated by 12 h, but half of them were ovulated by 24 h in response to FPH, whereas around 30% of oocytes were ovulated by 12 h and maximum ovulation (around 50%) occurred by 24 h in response to TPA. Without any stimulation (control), no ovulation occurred. TPA elevated the level of PGF$_{2a}$ to high levels when compared to control (basal levels), but the increase by FPH was less evident. Likewise, the levels of PGE$_2$ increased markedly in response to TPA, but rather decreased by FPH treatment. Interestingly, PGF$_{2a}$ induced ovulation but PGE$_2$ suppressed FPH- or PGF$_{2a}$-induced oocyte ovulation. Basal levels of PGs Increased steadily during culture. When theca/epithelium (THEP) layer and granulosa cell-enclosed oocytes (GCEOs) were separated by microdissection and cultured independently, higher levels of both PGs were secreted by THEP than by GCEOS. Synthesis of PGs by follicle or follicular components was strongly suppressed by exogenous cAMP or indomethacin. These results suggest that: 1) PGF$_{2a}$ plays an important role in Rana ovulation, 2) protein kinase C is involved in PGs production, and 3) thecal epithelium layer is responsible for the PGs production in Rana.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.12
• /
• pp.1827-1831
• /
• 2007

Effects of daidzein on expression of mRNAs of gonadotropin receptors (FSHR, LHR) and P450 aromatase (P450arom) were evaluated in ovarian follicles of white silky fowls. The hens were 13 months old in the post-peak period of egg laying and were randomly allocated as control and daidzein-treated groups, with daidzein supplemented to the basal diet at 10 mg/kg for 7 consecutive weeks. The mRNA expression of related genes was measured by semi-quantitative RT-PCR in the granulosa layers of the preovulatory follicle (PRF: F1, F2 ...) and follicular layers of the small yellow follicle (SYF), large white follicle (LWF) and atretic follicle (ATF). Results showed that daidzein supplementation significantly increased the number of SYF and LWF (p<0.05). The relative abundance of the FSHR mRNA decreased in the granulosa layers from F3 to F1, but LHR mRNA displayed opposite developmental changes. P450arom mRNA was highest in the SYF, but was very low in the granulosa layers after follicles finished selection. Treatment with daidzein resulted in increased mRNA expression of FSHR in F3 granulosa layer, LHR in granulosa layers of F3 to F1 and P450arom in LWF (p<0.05). These results indicated that dietary supplementation of daidzein up-regulated mRNA expression of gonadotropin receptors and P450arom to improve the development of preovulatory follicles in white silky fowls after the peak-laying period.

• Korean Journal of Veterinary Research
• /
• v.34 no.4
• /
• pp.873-880
• /
• 1994

Progesterone catabolizing enzyme, the enzyme $20{\alpha}$-hydroxysteroid deltydrgenase($20{\alpha}$-HSD) is pivotal in the regulation of ovarian function in rodents, which catabolizes progesterone into biologically inactive $20{\alpha}$ hydroxypregn-4-en-3-one($20{\alpha}$-OHP). In this study was carried out the influence of $20{\alpha}$-HSD activity to ovarian function and regulation such as ovulation, formation of corpus luteum, maintenance of estrous cycles and pregnancy, we investigated changes in activities of the splenic macrophages and ovarian histological findings in rat. During the estrous cycles, the ratious of phagocytotic macrophage in splenic adherent cells were highest on the proestrous, but they were lowest on metestrous. During the pregnancy, the ratious of phagocytotic macrophages in splenic adhrent cells were lowest by 1 day and then significantly increased toward the pregnancy, which were highest pregnancy on day 12. On histological findings in rat ovary, the mean number of growing, antral and mature follicles were 15.9(72.9%), 3.4(15.9%), and 2.4(11.2%), respectively. Growing follicles reached to lowest number at diestrus and mature follicle reached to lowest number at metestrus. The numbers of corpus luteums per tissue section of the ovary were 14.4 and the number of normal and atretic follicles were 11.0(76.4%) and 3.4(23.6%), respectively. The number of corpus luteum with vacolated cell were 1.7(11.8%). In this study suggests splenic macrophages are as a source of the substance which maintainer progesterone secretion from luteal cells, and are recognized pass the vessels and reside in the fresh corpora lutea soon after ovulation via inflammatory reactions, and these macrophages are felt to have a stimulatory effect on the formation of cropora lutea. In view of the results include previous report, ovarian $20{\alpha}$-hydroxysteroid dehydrogenase enzyme and splenic macrophages consider play central role in the control and maintenance of estrous cycles and pregnancy, and also applicable to both clinical and research in a wide variety such as control of reproductive system.

• The Korean Journal of Zoology
• /
• v.30 no.4
• /
• pp.351-370
• /
• 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

• Korean Journal of Clinical Laboratory Science
• /
• v.49 no.1
• /
• pp.40-47
• /
• 2017

Granulosa cells, which surround the oocyte within the ovarian follicle, play an essential role in creating conditions required for the development of oocytes and follicles. The solute carrier family (SLC) is comprised of influx transporters of steroidal hormones, various drugs, and several other substrates. The differential expression of selected DEGs was confirmed using in situ hybridization analysis. SLC23A3 and SLC39A10 were highly expressed in the ovary. The SLC39A10 gene was expressed in the primordial follicle stage, but SLC23A3 was expressed in the growing follicle stage. Contrastingly, the expression of SLC23A3 was increased in granulosa cells at the growing follicle stage. The differential expressions of SLC23A3 and SLC39A10 between the primordial and primary follicles were additionally confirmed by using follicle isolations. The gene expression profile from the present study may provide insight for future studies on the mechanism(s) involved in primordial-primary follicular transition and suggestions to promote follicular development in ovarian dysfunction.

• Clinical and Experimental Reproductive Medicine
• /
• v.47 no.3
• /
• pp.207-212
• /
• 2020

Objective: Glutamate ionotropic receptor AMPA type subunit 1 (GRIA1) is a subunit of a ligand-gated ion channel that regulates the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by controlling the release of gonadotropin-releasing hormone. Few studies have investigated the association between the GRIA1 gene and human infertility. This study evaluated the association of the GRIA1 rs548294 C > T and rs2195450 G > A polymorphisms with the ovarian response to human menopausal gonadotropin (HMG) in Iranian women. Methods: One hundred women with histories of at least 1 year of infertility were included. On the second day of menstruation, patients were injected with HMG; on the third day, blood samples were collected. After hormonal analysis, the GRIA1 rs548294 C > T and rs2195450 G > A genotypes of samples were identified via the restriction fragment length polymorphism method, and on day 9, the number of follicles was assessed via ultrasound. Results: For the GRIA1 rs548294 C > T and rs2195450 G > A single nucleotide polymorphisms, the subjects with CT and GG genotypes, respectively, displayed the highest mean FSH level, LH level, and number of follicles on day 9 of the menstrual cycle (p< 0.05). Significant positive correlations were observed between LH and FSH (p< 0.01), LH and follicle count (p< 0.01), FSH and age (p< 0.05), follicle count and age (p= 0.048), and FSH and follicle count (p< 0.01). Conclusion: This study showed a significant relationship between GRIA1 polymorphisms and ovarian response to the induction of ovulation. Therefore, determining patients' GRIA1 genotype may be useful for improving treatment and prescribing suitable doses of ovulation-stimulating drugs.

• Journal of Embryo Transfer
• /
• v.11 no.1
• /
• pp.41-50
• /
• 1996

This experiment was carried out to investigate the ovarian responses of the ovulation point, ovarian weight and size, the number of ovarian follicles and collected embryos, and to study the effects of the developmental stages (oocytes, 2-4 cell. 8-16 cell and morulae), additional levels of Ficoll (0, 15, 30%) on the survival rate (FDA-test) of rat embryos frozen in vitrification solution (20% glycerol + 10% ethylene glycol + 10% sucrose). Sunanarized results was as follows; 1. The mean ovulation point per head was 7, and the weight of ovaries was 0.03g. The size of ovary was 5.9 mm(L) and 4.6 mm(W), and the number of ovarian follicles over and below 2 mm was 4.7 and 8.7, respectively. The number of the collected embryos per head was 5.5 (79%). 2. 2. The FDA score of embryos frozen in 20 G 10 E 10 S without Ficoll was 2.8 (oocyte), 2.6 (2-4 cell), 3.9 (8-16 cell) and 3.6 (morula), respectively. However, there were no significant differences among treatments. 3. The FDA score of embryos frozen in 20 G 10 E 10 S with 15% Ficoll was 3.4 (oocyte), 4.0 (2-4 cell), 4.7 (8-16 cell) and 4.8 (morulae), respectively (P>0.05). 4. The FDA score of embryos frozen in 20 G 10 E 10 S with 30 % Ficoll was 3.7 (oocyte), 3.2 (2-4 cell), 4.4 (8-16 cell) and 4.4 (morulae), respectively (P>0.05). 5. As shown in the above results, the higher survival rate was obtained in the treatment of 15% Ficoll than that of 30%. And the survival rate (FDA-test)of the oocytes and 2-4 cell stages of the rat embryos was lower than that of 8~16 cell and morulae stages. It was considered that 8-16 cell and morulae could be available for the successful freezing by vitrification of rat embryos with 15% Ficoll except for oocytes.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.25 no.2
• /
• pp.66-77
• /
• 2012

Objectives: This study was designed to investigate the effects of PR on ovarian tissue in PCOS rats through measurement of morphological and histo-pathological observations, ovarian size. In addition, effects on expression levels of Insulin like Growth Factor Receptor(IGFR) were also investigated to elucidate related mechanisms. Methods: PCOS was induced by single intermuscular injection with ${\beta}$-Estradiol 17-Valerate(EV) in female rats. Normal group(NOR, n=8) were injected with sesame oil and administrated hard food for five weeks. Control group(CTL, n=8) were injected with EV and administrated hard food for five weeks. CR group(n=8) were injected with EV and administrated hard food mixed CR for five weeks. Then, we measured weights of body and ovary, uptakes of food and water. And we observed morphological and histo-pathological changes of ovary, levels of IGFR. Results: In this experiments, single injection of Estradiol Valerate(EV) induced suppression of weight gain, formation of cysts, increase of IGFR expression. Oral administration of PR prevent decrease of ovarian size significantly. Further more, formation of cystic follicles induced by EV injection is suppressed by PR treatment. Conclusions: These results suggest PR can be used for patients with PCOS to prevent formation of cystic follicles and malfunction of ovary.

• Journal of Radiation Protection and Research
• /
• v.23 no.2
• /
• pp.89-96
• /
• 1998

To assess the radioprotective effects of follicle stimulating hormone (FSH) on ovarian follicles, 3 week-old female mice were irradiated with 8.33 Gy of ${\gamma}$-ray (group R) and followed by 5 IU ip-injection of FSH (group RF). For control groups, 5 IU of saline (group C) or 5 IU of FSH (group F) was ip-injected. Ovaries were collected 0h, 6h, 12h, 14, 2d, 4d, and 8d after irradiation or saline/FSH injection, and followed by fixation in neutral buffered formalin for routine histochemistry. Immunohistochemistry was used to assess the status of follicles and DNA fragmentation was analyzed by agarose gel electrophoresis for total DNA. Staining specific for apoptotic follicles showed high intensity at 6h and 12h in group R and RF On the other hand, staining specific for proliferating follicles showed noticeably high intensity at 8d in group R and Rf. DNA fragmentation of 185bp increased with time in all experimental groups. Especially 370bp appeared at 6h in group R, then disappeared after 1d. In case of group RF, it appeared at 12h and disappeared after 1d. From the above results, the irradiated antral follicles become completely disappeared from 4d to 8d, and then new follicles started to grow again at 8d. FSH had delaying or suppressing effects on follicular atresia after irradiation. In addition, it became clear that radiation-induced follicular atresia was mediated by granulosa cell apoptosis.