• Animal cells and systems
• /
• v.1 no.4
• /
• pp.595-602
• /
• 1997

Treatment of newborn female rats with gonadal steroids induces permanent sterility in adulthood. We investigated the alteration in expression patterns of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) in neonatally estrogenized sterile rats (ESR). Newborn female rats received daily injections of 17${\beta}$-estradiol (E, 10 ${\mu}$g) from the day of birth (day 1) to postnatal day 5. Controls were subjected to vehicles over the same period. All animals were sacrificed on week 7 after birth. Hypothalamic GnRH mANA levels were markedly higher in all ESR than in controls, while hypothalamic GnRH contents in ESR increased in proportion to the frequency of daily administration of E. However, both pituitary LH6 mRNA and serum LH levels were inversely decreased by the same treatment. The data indicate that neonatal exposure of E equally elevates the expression of GnRH gene, but reduces the secretion of GnRH, accordingly leading to attenuation of LH6 gene expression and circulating LH levels. The temporal effect of E and/or progesterone (P) on GnRH and LH6 mRNA levels was also examined in ESR. Newborn female rats were daily injected with E (10 ${\mu}$g) or vehicle for five successive days from day 1 and ovariectomized at week 5. They were implanted with E (235 ${\mu}$g/ml) two days prior to week 7, injected with P (1 mg) 42 h later, and sacrificed 7 h after P administration. In ovariectomized controls, hypothalamic GnRH mRNA levels were dropped to half by treatment of E and restored by subsequent treatment of P. The negative feedback action of E on GnRH mRNA levels observed in ovariectomized rats was completely blocked by neonatal exposure of E. The change in pituitary LH mRNA levels was similar to that in hypothalamic GnRH mRNA levels. Taken together, the results suggest that neonatal treatment of E alters the synthesis and release of GnRH in adulthood and furthermore blocks the negative feedback regulation of E which occurs normally after ovariectomy.

• Development and Reproduction
• /
• v.4 no.2
• /
• pp.187-193
• /
• 2000

Gonad and blood samples were taken from the cultured female Korean sea bass, Lateolabrax japonicus from October to February between 1997 and 1999. Gonadosomatic index began to increase in November and reached the highest value in December (12.8$\pm$1.5) and January (14.8$\pm$3.5), and then decreased sharply in February (2.6$\pm$1.5, p<0.05). The ovarian oocytes developed to tertiary yolk stage and reached fully-Brown stage in December and January, and then underwent atresia without maturation and ovulation in February. The plasma estradio3-17 $\beta$ level increased from November, and reached the highest value in December (1,152.3$\pm$107.2 pg/ml) and January (1,315.4$\pm$99.5 pg/ml), after then decreased in February (P<0.05). The concentration of plasma 17 $\alpha$ ,20 $\beta$-dihydroxy-4-pregnen-3-one was not significantly changed at low levels (86.6$\pm$6.5∼93.8$\pm$2.8 pg/ml) during the experimental period (P<0.05). All the fish with fully-grown oocytes in the ovary were matured and ovulated by HCG injection. The number of floating eggs were 325,000$\pm$26,000 at HCG 1,000 luhg and 195,000$\pm$35,000 at 2,000 lUikg. There was no difference in fertilization rate and hatching rate of the eggs (P<0.05). Considering these results, we could infer that the ovarian oocyte of the cultured Korean sea bass were not matured and ovulated because of the lack of gonadotropin surge. Moreover, HCG injection could induce oocyte maturation and ovulation in the cultured fish, and the effective dose was 1,000 IU/kg.

• Korean Journal of Veterinary Service
• /
• v.23 no.3
• /
• pp.247-254
• /
• 2000

This study was performed to elucidate the effects of antler, red ginseng, safflower seed, ipriflavone and estrogen on ovariectomized rats. The rats were fed with Ca and P deficient diet for five weeks to induce osteoporosis. After this period, these animals were fed with normal feed and treated every other day with antler(600 mg/kg, p.o), red ginseng(200 mg/kg, p.o), safflower(200 mg/kg, p.o), ipriflavone(80 mg/kg, p.o) and estrogene(400$\mu\textrm{g}$/kg, i.m) for 5 weeks. During the treatment, the rats were examined for serum concentrations of estradiol, calcitonin, Ca, p and alkaline phosphatase(ALP) activities. The results are summarized as follows 1. The levels of serum 17 $\beta$-estradiol after five weeks of treatment were showed 39.6$\pm$3.0 pg/ml by antler, 33.2$\pm$2.5pg/ml by red ginseng, 34.9$\pm$2.4pg/ml by safflower, 28.1$\pm$3.1 pg/ml by ipriflavone and 40.6$\pm$3.0pg/ml by estrogen-treated group. They were lower than 50.8$\pm$3.1pg/ml of normal control group which had not received ovariectomy. They, however, were significantly higher than 26.8$\pm$1.8pg/ml of ovariectomized non-treatment group(p<0.05). 2. The levels of serum calcitonin after five weeks of treatment were showed 0.60$\pm$0.02 ng/ml by antler, 0.55$\pm$0.04ng/ml by red ginseng, 0.59$\pm$0.02ng/ml by safflower, 0.56$\pm$0.04ng/ml by ipriflavone and 0.62$\pm$0.02ng/ml by estrogen-treated group. They were lower than 0.67$\pm$0.03pg/ml of normal control group. However, they were significantly higher than 0.45$\pm$0.05ng/ml of ovariectomized non-treatment group(p<0.05). 3. The levels of serum Ca of the rats after five weeks of treatment with antler, red ginseng, safflower, ipriflavone and estrogen were 23.51$\pm$2.19$\mu\textrm{g}$/$m\ell$, 25.22$\pm$3.44$\mu\textrm{g}$/$m\ell$, 23.20$\pm$0.02$\mu\textrm{g}$/$m\ell$, 24.76$\pm$3.57$\mu\textrm{g}$/$m\ell$, 23.07$\pm$3.66$\mu\textrm{g}$/$m\ell$, respectively. They were a bit higher than 21.43$\pm$2.22$\mu\textrm{g}$/$m\ell$ of normal control group. And non-treatment group showed 26.12$\pm$0.29$\mu\textrm{g}$/$m\ell$ which was significantly higher than that of control group(P<0.05). 4. The serum P concentraions after five weeks of treatment were showed 12.11$\pm$2.14$\mu\textrm{g}$/$m\ell$ by antler, 13.18$\pm$1.64u91m4 by red ginseng, 12.67 $\pm$2.31$\mu\textrm{g}$/$m\ell$ by safflower, 12.38$\pm$2.07$\mu\textrm{g}$/$m\ell$ by ipriflavone, 11.86$\pm$1.93$\mu\textrm{g}$/$m\ell$ by estrogen-treated group. They were a bit higher than 11.29$\pm$1.23$\mu\textrm{g}$/$m\ell$ of normal control group. And non-treatment group showed 13.42$\pm$1.87$\mu\textrm{g}$/$m\ell$ which was higher than that of control group but not significant. 5. The levels of serum ALP after five weeks of treatment were showed 164.8$\pm$3.8IU/ml by antler, 277.7$\pm$4.8IU/ml by red ginseng, 288.5$\pm$4.5IU/ml by safflower, 214.7$\pm$5.7IU/ml by ipriflavone and 159.4$\pm$5.4IU/ml by estrogen-treated group. They were significantly higher than 144.1$\pm$3.5IU/ml of normal control group(p<0.05). However, they were significantly lower than 336.9 $\pm$12.7IU/ml of ovariectomized non-treatment group(p<0.05). Antler and safflower elevated serum estradiol and calcitonin, and decreased serum ALP significantly. Therefore they were thought to have therapeutic effect on osteoposis by making inhibitory effect on osteoclasts rather than activating osteoblasts.

• Journal of Embryo Transfer
• /
• v.22 no.1
• /
• pp.21-26
• /
• 2007

본 연구는 mSOF(modified synthetic oviduct fluid medium) 배양액을 이용하여 $100{\mu}1$와 $10{\mu}1$ 배양 소적에서 일본 흑우의 수정란 생산 효율을 개선하기 위하여 수행하였다. 난구세포가 부착된 미성숙 난자는 각각 단독 배양조건($S;\;10{\mu}1$ 소적) 및 그룹 배양 조건 ($G;\;100{\mu}1$ 소적)에서 실시하였고 배양액은 TCM-199의 기본 배지에 10% FCS, 0.02IU/ml FSH와 $1{\mu}g/ml$ $estradiol-17{\beta}$를 첨가하여 사용하였다. 배반포 단계로 발육한 수정란은 1.5M ethylene glycol로 직접 이식법에 의한 동결 방법으로 동결을 실시하였고, 세포수는 융해 후 생존 수정란에 대해 조사하였다. 체외 배양 시간이 $16{\sim}17$시간 배양 조건에서 난자의 성숙율은 그룹 배양 조건$(27.1{\pm}16.8%)$보다는 단독 배양조건$(57.1{\pm}15.0%)$에서 성숙율이 높았다(p<0.05). 그러나 체외 배양 시간이 $18{\sim}19$ 시간과 $20{\sim}21$시간 배양시는 유사한 성숙율을 보였다. 난자의 체외 배양율은 체외 배양 시간의 증가에 의해 성숙도가 $86.3{\pm}9.9%$로 증가하였다. 접합체(zygote)의 분할율은 단독이나 그룹 배양 조건에서도 유사한 결과를 얻었다. 배반포 발달율은 배양 $7{\sim}8$일째에 조사한 결과 단독 배양 방법보다는 그룹 배양 방법에서 발달율이 높았으나, 분할된 접합체를 기준으로 한 경우 배반포 발달율$(S;\; 21.4{\pm}10.6%,\;G;\;39.0{\pm}13.1%)$에서는 유의적인 차이가 없었다. 단독 배양과 그룹 배양에서 $6.5{\pm}8$일 사이에 배반포로 발달된 수정란의 세포수 조사에서는 유사한 결과를 얻었다. 동결 융해 후 24시간 배양 후 배반포 생존율(5; 24.2%, G; 30.2%), 부화율(S: 20.9%, G: 12.7%) 및 생존 수정란수(S; 45.2%, G: 42.8%)에서도 배양 조건에 따른 유의적인 차는 없었다. 결론적으로 mSOF배양액을 이용하는 경우 미성숙 난자의 체외 성숙 유도 배양 시 단독이나 그룹 배양 시 배반포 발달율에서 그룹간에 유의적인 차가 인정되었다(p<0.01).

• Journal of Food Hygiene and Safety
• /
• v.21 no.2
• /
• pp.100-106
• /
• 2006

The use of underarm and body care cosmetics with oestrogenic chemical excipients (particularly the parabens) and the hypothesized association with breast cancer incidence, particularly in women. It is noted that the type of cosmetic product is irrelevant (e.g. antiperspirant/deodorant versus body lotion, moisturizers or sprays versus creams) and attention must focus on issues of actual exposure to chemicals through continued dermal application of body care products and the endocrine/hormonal activity and toxicity of the chemicals in the formulations. To evaluate the estrogenic activities of parabens such as ethylparaben, butylparaben, propylparaben, isobutylparaben and isopropylparaben, we used recombinant yeasts containing the human estrogen receptor [Saccharomyces cerevisiae ER+LYS 8127], human breast cancer MCF-7 cell lines and human estrogen receptor ${\alpha}\;and\;{\beta}$. In E-screen assays, isopropylparaben is the most estrogenic paraben, and in ER competition assay, isobutylparaben is the most estrogenic paraben. We evaluated isopropylparaben was most active in the recombinant yeast assay, followed by propylparaben, ethylparaben, isobutylparaben and butylparaben. Results from this study demonstrate that parabens are observed in human endocrine system. Therefore, we have shown that the parabens is induced the estrogenic activities similar to $17{\beta}$-estradiol and Bisphenol-A.

• Korean Journal of Agricultural Science
• /
• v.9 no.1
• /
• pp.303-313
• /
• 1982

The study was conducted to find out the concentrations of sex hormones and the contents of metabolites in serum, and the changes of weights and tissues on ovary, thyroid gland and adrenal gland according to gestation period in rabbit. The results were summarized as follows: 1. The ovary weights were increased significantly with the time e lapse after gestation and recovered normally at 5 days after parturition. In the histological changes of ovary, the lutein cells were hypertrophied and the secretory granules were increased actively until 3 weeks after gestation, and then a trophied thereafter. 2. The thyroid gland weights at all observation times were higher than those in control group, and the significance was recognized at 1, 3 and 4 weeks after gestation. The histological features of the secretory epithelium were the hyper trophic and columnar condition stimulating the functional state from 1 week after gestation. 3. The adrenal gland weights in experimental group were recognized significantly at 4 weeks after gestation, but showed higher than those in control group at all observation times. The zona fasciculata and zona reticularis of the gland showed the slight hypertrophic condition, but the zona glomeerulosa and adrenal medulla did not find out any particular changes. 4. The serum concentrations of progesterone and LH reached a peak level at 2 weeks and 1 week after geestation respectively, and rapidly began to decline thereafter. 5. The serum concentrations of estradiol-$17{\beta}$ and FSH were not detected below 20.0 pg/ml and 1.3 mIU/ml respectively. 6. The contents of total protein and non-protein nitrogen nitrogen were decreased gradually with the time elapse after gestation, but the significant differences were recognized from 3 weeks. 7. The total lipids were not changed markedly until 3 weeks, but increased significantly at 4 weeks after gestation and at 5 days after parturition. 8. The serum cholesterol tended to be decreased until 3 weeks, but increased at 4 weeks after gestation and at 5 days after parturition. 9. The serum calcium showed a continuous decrease during the gestation period, but the significant differences were recognized at 3 and 4 weeks. The serum phosphorus also had a significant decrease at 4 weeks after gestation.

• Journal of Aquaculture
• /
• v.21 no.4
• /
• pp.285-293
• /
• 2008

Vitellogenin (Vg) is the precursor of vitellin (Vn), the major yolk protein of teleost fishes. In this study, Vg and Vn proteins of the Korean bullhead Pseudobagrus fulvidraco were isolated using gel-filtration chromatography (Sephadex-G 200 column) and anion-exchange chromatography (Mono Q HR 5/5 column), respectively. Purified Vn with an estimated molecular mass of 360 kDa by gel filtration chromatography was obtained from ovarian egg, and it was composited to one major subunit with an estimated molecular mass of 107 kDa by SDS-PAGE. In the result of western blotting, one major band was detected using antiserum against Vn (anti-Vn). These results suggested that Vn was composed of three subunits having the same molecular weight in Pseudobagrus fulvidraco. Vg was induced by estradiol-$17{\beta}$ ($E_2$) and purified from $E_2$ treated male serum. The molecular weight of whole Vg was estimated to be 450 kDa by gel filtration chromatography, and it is composed of three subunits with estimated molecular masses of 110 kDa, 125 kDa and 147 kDa as determined by SDS-PAGE. In the Ouchterlony's immunodiffusion test using anti-Vn and antiserum against female and male serum, purified Vg was detected in matured female and Ez treated male serum but not in untreated male. These results can be used in detecting estrogenic contamination of the aquatic environment.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.34 no.1
• /
• pp.13-16
• /
• 2001

Plasma levels of sex steroid hormones in rockfish, Sebastes inermis were examined monthly in relation to gonadosomatic index (GSI) under a controlled water temperature and photoperiod, The GSI of a control group (C) in female began to increase from November and reached a maximum in January, Sample fish under a controlled water temperature and photoperiod (Tr) were divided into a responded group (Tr-r) and a un-responded group (Tr-n) by the gonadal maturation condition and GSI. The GSI of females in Tr-r reached a maximum in March. But the female GSI in Tr-n kept lower than 1.2 during the experimental period. No differences in male GSI were noticed between C and Tr. The $estradiol-17\beta$ and testosterone levels of female plasma in Tr reached a maximum in October, later than those in C. In males, these was no difference in 11-ketotestosterone and testosterone between C and Tr. When rockfish was reared in September under the controlled water temperature and photoperiod which were equivalent to those in July, that is two months earlier, the maturation of females was delayed in comparison with C. This finding suggested that delayed maturation in ovary was caused by the secretion of sex steroid hormones in relation to the water temperature and photoperiod of the hypothalamus-pituitary-gonad axis.

• Journal of Embryo Transfer
• /
• v.24 no.3
• /
• pp.199-205
• /
• 2009

This study was carried out to investigate the effective genetic resources preservation system using the frozen boar semen. The porcine oocytes were matured for 44 hours in NCSU-23 medium with or without 10% Porcine Follicle Fluid (PFF), 0.5 ${\mu}g/ml$ porcine FSH, 0.5 ${\mu}g/ml$ equine LH, 1.0 ${\mu}g/ml$ 17 $\beta$-estradiol ($E_2$) and 10 ng/ml Epidermal Growth Factor (EGF) under mineral oil at $38.5^{\circ}C$ in humidified atmosphere of 5% $CO_2$ in air. After 44 h of culture, the oocytes were inseminated with frozen-thawed semen and fresh semen prepared with mTBM medium for 6 h. Later, set of 50 presumptive zygotes were transferred into 4-well dish (500 ${\mu}l$) of IVC medium. for embryos freezing, slow-freezing and vitrification methods were used as a cryopreservation. Differences among treatments were analyzed using General Linear Model Procedure by SAS Package (version 6.12) differences were considered significant when p<0.05. Following IVF and IVC, the rates of cleavage and blastocysts formation were significantly higher (p<0.05) in hormone supplemented group than that of hormone-free group (25.7 vs, 12.1). The development rates to cleavage and blastocysts were significantly higher in PZM-5 group than NCSU-23 group (60.3%, 46.6% vs 27.4%, 11.1%). Further improvement was achieved when PZM-5 was supplemented with FBS. Cleavage rates was significantly higher in fresh semen source group than frozen semen (66.7% vs 43.7%). However in blastocysts rates was similar two groups. Post-thaw survival rates of embryos were 1.2% and 2.2% in slow-frezing and vitrification groups, respectively. The results of our study suggest that it is still possible to improve the culture conditions and boar semen cryopreservation for enhance reproductive technology and animal genetic resources conservation.

• Journal of Embryo Transfer
• /
• v.29 no.1
• /
• pp.35-41
• /
• 2014

Laparoscopic ovum pick-up (LOPU) is a convenient method for collecting oocytes in small ruminants. LOPU has the advantage of being a less invasive means of oocyte collection, thereby allowing for a repeated usage of the oocyte donor animals. A total of 25 Korean black goats were used in the winter season (December to February) and LOPU was applied to the goats which had been treated for superovulation more than two times during the last twelve months. Estrus was synchronized with an intravaginal insert containing 0.3 g progesterone for 10 to 12 days. Ovaries were hyperstimulated with eCG 1,000 IU oneshot, FSH with eCG (50 mg / 1,000 IU; 70 mg / 500 IU; 70 mg / 1,000 IU) oneshot or FSH multiple-shot with eCG oneshot ($20mg{\times}6/300IU$) given intramuscularly 72 h prior to LOPU. For these groups, the number of follicles (mean ${\pm}$ SEM) observed which developed to larger than 2 mm in diameter were $1.6{\pm}2.5$, $4.3{\pm}3.1$, $5.5{\pm}4.2$, $6.6{\pm}2.1$ and $8.8{\pm}7.8$, respectively. Oocytes were aspirated by using OPU needles and a vacuum pump. The overall oocyte retrieval rates were 41.4%. Oocytes were matured in TCM-199 supplemented with 10% (w/v) bovine serum albumin + $10{\mu}g/ml$ FSH + $1{\mu}g/ml$ $17{\beta}$-estradiol for 27 h at $39^{\circ}C$ in 5% $CO_2$ in air. Oocytes were parthenogenetically activated by ionomycin combined with 6-diethylaminopurine (6-DMAP). Total oocyte maturation and cleavage rate were 67.3% and 78.8%, respectively. In summary, LOPU is a useful oocyte collection method in Korean black goats that can provide immature oocytes for transgenesis or nuclear transfer.