• Clinical and Experimental Reproductive Medicine
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• v.33 no.1
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• pp.35-35
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• 2006

Objective: The purposes of the present study were to investigate the effect of ${\gamma}$-radiation on the expression of inhibin-${\alpha}$ proteins and genes for inhibin ${\alpha}$, ${\beta}A$, and ${\beta}B$ in the ovary. Methods: Immature mice were whole-body ${\gamma}$-irradiated with 25% of a lethal dose. At time 0, 3, 6, 12, and 24 hours after the irradiation, the ovaries were collected and used for immunohistochemistry for inhibin-${\alpha}$, and RT_PCR for inhibin-${\alpha}$, ${\beta}A$, and ${\beta}B$. Results: The expression of the immunoreactive inhibins-${\alpha}$ was maintained at 12 hours post-irradiation and reduced thereafter. The expression of inhibin-${\alpha}$ mRNA was significantly increased with the time after the irradiation. However there were no significant changes in the expression of ${\beta}A$ and ${\beta}B$ mRNAs. Conclusion: It might be thought that inhibin acts as one of the regulatory factors in the ${\gamma}$-radiation-induced follicular atresia in mice

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.119-124
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• 2001

This study was performed to investigate the effects of the peptide to carrier ratio on the immune and biological functions to inhibin immunization in Hanwoo. A peptide sequence kom the alpha -subunit (19~32 peptide) of porcine inhibin was synthesized for antigen and conjugated to human serum albumin(HSA) for carrier protein. Anti-inhibin sera(AI) were produced 52 day later from rabbit after injection of inhibin-$\alpha$ -subunit peptide conjugator for antigen with the interval of 2 weeks. Immune-blotting analysis using antibody specific fur inhibin-$\alpha$ subunits revealed that the inhibin was detected at 1.0 cm bovine follicular fluid(bFF). However, each stage of corpus lutea and 0.1 cm of follicular fluid were not detected. The maximal contents of estradiol-17 $\beta$ in Hanwoo ovarian follicular fluid were detected at 2.0 cm of follicular size(diameter), but the mean total contents of these hormone decreased significantly with decreasing diameter of follicles. However, progesterone contents of follicular fluid were high at 1.0 cm of follicle. Progesterone secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p＜0.05) inhibited in 5% bFF and 5% bFF + 5% AI addition group compared with control group. Estradiol-17 $\beta$ secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p＜0.05) increased in 5% AI and 5% AI + 5% bFF addtion group compared with control group. However, the groups added 5% AI were not changed compared to control groups in progesterone and estradiol-17 $\beta$. Taken together, we suggested that inhibin in the mature FF plays a pivotal role on the biosynthesis of steroid hormone of follicular cells during follicular development.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.34-34
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• 2001

본 연구의 목적은 한우 난포발달에 있어서 난포액 및 anti-inhibin serum의 생리적 역할을 검토하기 위해 수행하였다. Anti-inhibin serum(AI)은 항원으로서 porcine inhibin-$\alpha$-subunit 19~32의 peptide를 사용하여 adjuvant 용액을 1:3의 비율로 혼합하여 앙고라종 토끼 5두(체중 2.5kg)에게 주 2회 간격으로 접종 후 얻어진 항혈청을 사용하였다. 난포액(bFF; bovine follicular fluid)은 도축장에서 도축되는 한우 난소로부터 직경 1.0cm 이하의 난포로부터 회수하여 스테로이드를 제거하기 위해 10% chacoal so lution(50 mg/$m\ell$, Norrit-A, Fisher Sci., USA)을 처리하여 45분간 배양후 원심분리후 상층액을 회수하여 실험에 공시하였다. 공시동물은 1산후 정상적으로 발정주기가 반복되는 한우암소 9두를 난소감정후 황체를 확인하여 PGF2$\alpha$제제(lutylase. USA)를 주사하여 발정을 유기한 다음, 난소의 first wave가 시작되는 시기인 배란직후 12시간째부터 4일간 일일 2회 5 $m\ell$씩 총 8회 40 $m\ell$의 AI와 bFF를 각각 경정맥으로 주사하였으며 대조구로서 생리식염수를 주사하였으며 채혈 및 정맥주사를 용이하게 하기 위해 경정맥에 카테타를 설치하여 6시간간격으로 총 200시간까지 채혈하였으며 초음파진단기를 이용하여 난포의 발달을 검토하였다. 채혈후 혈중Inhibin, progesterone 및 Estrad iol-17$\beta$농도의 분석은 RIA 및 ELISA법으로 분석하여 얻어진 결과는 다음과 같다. 혈중 Progesterone농도는 대조구와 AI처리구에서는 배란후 68시간째부터 유의적으로 증가하기 시작하였으나, bFF처리구에서는 배란후 68시간부터 170시간까지 대조구에 비해 유의적으로 낮은 농도를 나타내었다. 이에 반해 혈중 Estradiol-17$\beta$농도는 대조구의 경우 bFF처리구와 비슷한 수준으로 배란후부터 낮은 농도를 유지하였으나, AI 처리구는 배란후 36시간이후부터 108시간까지 유의적으로 높은 수준을 유지하였다가 그 이후 감소되었다. 한편, 혈중 Inhibin농도는 전 구간에서 배란후 84시간까지 불규칙한 농도를 보이다가 bFF처리구에서는 배란후 84시간부터 유의적으로 증가하였다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.33-33
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• 2001

본 연구는 한우 난포발달에 있어서 난포액 및 inhibin의 생리적 역할을 검토하기 위해 수행하였다. Anti-inhibin serum(AIS) 생산을 위해 사용된 항원은 porcine inhibin-$\alpha$-subunit 19~32의 peptide를 사용하여 adjuvant 용액을 1:3의 비율로 혼합하여 앙고라종 토끼 5두(체중 2.5kg)에게 주 2회 간격으로 8회 실시후 ELISA Leader로 항혈청의 역가를 확인하였다. 난포액(bFF; bovine follicular fluid)은 도축장에서 도축되는 한우 난소로부터 직경 1.0cm 이하의 난포로부터 회수하여 스테로이드를 제거하기 위해 10% chacoal solution(50 mg/$m\ell$, Norrit-A, Fisher Sci., USA)을 처리하여 45분간 배양후 원심분리후 상층액을 회수하여 실험에 공시하였다. 과립막세포의 체외배양을 위해 D-MEM용액(10% FCS와 antibiotics를 첨가)을 배양액으로 하여 1 $\times$ $10^{6}$ cells/$m\ell$로 조절하였다. 호르몬은 RIA 및 ELISA법으로 분석하였다. 그 결과 항원-항체반응은 항원처리후 24일째부터 항체가를 확인할 수 있었으며 52일째에서 높은 항원-항체반응을 보였다. 한편, 난포크기별 난포액의 progesterone 및 Estradiol-l7$\beta$을 농도를 분석한 결과, Progesterone 난포크기가 직경 1.0 cm부터 유의적으로 증가하기 시작하여 직경 2.0cm의 난포액에서는 높은 progesterone이 존재하고 있는 것으로 나타났다. 이에 반해, 난포크기별 난포액중 estradiol 17$\beta$농도는 직경 1.0cm구에서 가장 높게 나타났다. 난포직경이 1cm일 경우에 난포액내 etradiol-17$\beta$가 가장 많이 존재하고 있음이 나타났다. 과립막세포의 체외배양에서 배양 24시간에서는 과립막세포의 progesterone분비는 약 40ng/1$\times$$10^{6}$ cell/well/$m\ell$ 전후로 나타났으며 bFF 5%, AI 5% 및 bFF＋AI 첨가구에 따라 유의적인 차이는 없었다 반면에 48시간배양구에서는 24시간에 비해 유의적으로 높게 분비되었으며, bFF 5%처리구와 bFF5%＋AI5%처리구에서는 progesterone을 대조구보다 유의적으로 억제되었으나 AI 5%단독처리구에서는 대조구와 큰 차이가 없었다. 한편, 각 세포질을 SDS-PAGE로 분리하여 nitro cellulose membrane에 transfer하여 Western blotting법에 의해 검토한 결과, 직경 1.0 cm의 성숙난포의 granulosa cell에 특이하게 Inhibin이 존재하고 있음이 확인되었다. 이상의 결과로, 한우에 있어서 성숙난포에 존재하는 Inhibin은 난포발달 및 난포세포의 스테로이드호르몬합성에 중요하게 관여하고 있는 것으로 사료된다.

• Journal of Radiation Protection and Research
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• v.32 no.4
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• pp.140-156
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• 2007

This research was performed to investigate the morphological changes of folliculus ovary according to the radiation dose. The whole body radiation of 200 cGy, 400 cGy, and 600 cGy was given to the each groups of 5 months-aged female mouse. Various staining methods used in this research are: Hematosylin-Eosin method, and immunohistochemistrical methods using BrdU, TUNEL, p53, p21, PCNA and inhibin. The minute structural changes of folliculus ovary were observed through an electron microscope with high magnification. The morphological changes of growing folliculus ovary became distinct as the dose of X-rays increased. Especially, the nuclei of granular cells showed manifest condensation and the changes of the transparent zone were distinct. As a result of histochemical reaction according to Masson's trichrome method and reticular fiber method, the changed granular cells, the deformed basilar membrane of folliculus ovary and the abnormal arrangement of the reticular fiber were observed. In the reaction of BrdU, the granular cells of normal folliculus ovary with positive reaction rapidly decreased according to the increase of the dose of X-rays. In TUNEL study, granular cells showing positive reaction in retarded folliculus ovary were expanded to growing folliculus ovary and primordial folliculus ovary according to the increase of the dose of X-rays. In case of 600 cGy of X-rays, oocyte underwent apoptosis. In p53 immunohistochemistry, p53 manifested to be stronger as the dose of X-rays increased. p53 reactivity was manifested distinctively in all cells comprising folliculus ovary following irradiation of 600 cGy. p21 was manifested in granular cells of folliculus ovary and showed very positive reaction around follicular antrum according to the increase of the dose of X-rays. In PCNA, positive reaction was manifested in growing folliculus ovary, mature folliculus ovary and primordial folliculus ovary, but the extent of the reaction decreased as the dose of the X-rays decreased. The finding that the reaction of granular cells around folliculus ovary was stronger than that near follicular membrane indicates that what was damaged first by X-ray was the cells near folliculus ovary and follicular antrum. The reactivity of $inhibin-{\alpha}$ showed difference according to the growing stage of folliculus ovary: $inhibin-{\alpha}$ showed the most strong reaction in mature folliculus ovary with follicular antrum. There was strong reaction in granular cells around follicular membrane but $inhibin-{\alpha}$ did not occur at all in theca cells comprising follicular membrane. $Inhibin-{\alpha}$ in ovary tissue exposed to 400 cGy of X-rays was manifested more strongly than in ovary tissue exposed to 600 cGy of X-rays, which was related to the phenomenon that granular cells of mature folliculus ovary underwent necrosis or apoptosis increasingly due to X-rays. In an electron microscope with high magnification, nuclei and protoplasm of granular cells in growing folliculus ovary abruptly underwent minute structural changes according to the increase of dose of X-rays. Cell residue, by-product of cell decease, neutrophil and macrophage around follicular antrum were observed. The minute structural changes in granular cells showed typical characteristics of apoptosis: the increase of electronic density due to nuclear condensation, fragmentation of nuclei and atrophy of protoplasm. Necrosis of cells was identified but it was not so remarkable. Macrophage with apoptotic bodies was scattered. Proportional to the radiation dose, we found that the generation of heterogeneous substance of normal ovary texture's follicular fluid, the emergence of dyeing characteristic in the basilar membrane of folicle, the generation of apoptosis, and the transformation of macrophages, etc. From this results, we can infer the possible radiation hazard on the ovary of cervix cancer patient with radiation therapy.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.9
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• pp.1205-1209
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• 2000

The present study was undertaken to investigate the effect of stimulation of follicular development with eCG on the peripheral levels of inhibin and FSH in Murrah buffaloes. Estrus was synchronized in five normally cycling females by insertion of Crestar (Intervet, Boxmeer, Holland) implants for nine days. Estradiol valerate was administered i.m. on the day of implant insertion. On the 10th day of the induced estrous cycle a single dose of 3000 IU eCG (Folligon, Intervet, Boxmeer, Holland) was given, followed by treatment with 25 mg of $PGF_2$ alpha (Lutalyse, Upjohn, Belgium) 48 h later. Blood samples were obtained during the induced estrus, on cycle day 10 (luteal phase), at the superovulatory estrus (43 h after PGF) and during the periovulatory period (64 h after PGF). Ultrasonography was done daily to monitor follicular development. Plasma concentrations of inhibin and FSH were determined by specific radioimmunoassays. Differences between $mean{\pm}SEM$ values of different phases of the cycle were compared by ANOVA. The mean number of small (2-5 mm), medium (6-9 mm) and large (>10 mm) follicles observed two days after eCG treatment and on the day of superovulatory estrus was $2.8{\pm}0.31$, $5.2{\pm}0.30$ and $1.4{\pm}0.09$ and $1.9{\pm}0.21$, $2.8{\pm}0.40$ and $5.0{\pm}0.83$, respectively. The mean number of ovulations was $3.6{\pm}0.37$ and the mean number of unovulated follicles was $6.1{\pm}0.47$. Most of the follicles >10 mm in diameter had ovulated (72%). The mean ${\pm}SEM $ of plasma inhibin concentration $(2584.15{\pm}17.92pg/ml)$ during the superovulatory estrus was significantly higher $(p{\leq}0.05)$ than during the induced estrus $(749.87{\pm}17.29pg/ml)$, the luteal phase $(1099.54{\pm}24.98pg/ml)$ and periovulatory period $(1682.71{\pm}29.88pg/ml)$, respectively. $Mean{\pm}SEM$ plasma FSH concentration during the induced estrus $(10.35{\pm}0.41ng/ml)$ was not different from that during the superovulatory estrus $(8.52{\pm}0.39ng/ml)$, but was significantly higher $(p{\leq}0.05)$ than during the luteal phase $(2.81{\pm}0.42ng/ml)$ and periovulatory period $(5.7{\pm}0.28ng/ml)$. These data indicate that treatment with eCG in buffaloes for inducing superovulation results in a significant elevation in plasma inhibin levels and a decrease in plasma FSH levels during the superovulatory estrus. Thus, we suggest that the elevated plasma inhibin coming from fully developed follicles continued for a long time which results in inhibition of FSH leading to poor ovulation in the remaining follicles, which may be the cause of suboptimal superovulatory response.

• Journal of radiological science and technology
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• v.39 no.4
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• pp.549-555
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• 2016

This study was an evaluation of the significance of each parameter through aimed at pregnant women subjected to screening test(integrated test) in predicting risk of Down syndrome. We retrospectively analysed the correlation of risk of Down's syndrome with Nuchal Translucency(NT) images measured by ultrasound, Pregnancy Associated Plasma Protein A(PAPP-A), alpha-fetoprotein(AFP), unconjugated estriol(uE3), human chorionic gonadotrophin(hCG) and Inhibin A by maternal serum. As a result, a significant correlation with NT, uE3, hCG, Inhibin A is revealed with Down's syndrome risk(P<.001). In ROC analysis, AUC of Inhibin A is analysed as the biggest predictor of Down's syndrome(0.859). And the criterion for cut-off was inhibin A 1.4 MoM(sensitivity 81.8%, specificity 75.9%). In conclusion, Inhibin A was the most useful in parameters to predict Down's syndrome in the integrated test. If we make up for the weakness based on the cut-off value of parameters they will be able to be used as an independent indicator in the risk of Down's syndrome screening.

• Korean Journal of Veterinary Service
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• v.34 no.3
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• pp.273-278
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• 2011

Sertoli cell tumor (SCT) of the testicle arises from the supporting cells within the seminiferous tubules. SCT is common in dogs, especially in cryptorchid testicles, but also has been reported in the stallion, ram, cat, and bull. Sertoli cell tumor sample was collected from 7-years male german shepherd. In this study, SCT arose from one testicle. Sample size is approximately 1.7 cm in diameter and it has a round form. In the microscopic, cells within the tumor variably resemble Sertoli cells (SCs) that normally populate the seminiferous tubules and interstitial area. There is abundant stroma of dense, mature fibrous connective tissue in SCT. In the immunohistochemical staining, cytokeratin AE1/AE3 was not expressed in the control and SCT. S-100 protein was expressed by SCs, germ cells and fibrous connective tissue of SCT. Melan A was expressed by leydig cells (LCs) of SCT. A study by using S-100 and melan A in canine SCT was almost never carried out. S-100 and melans A is considered to suggest for diagnosis and pathogenesis of canine SCTs. Inhibin-alpha and Vimentin were well known as the marekers of SCTs. Also, they were expressed by Sertoli cells and LSs of SCT in this study.

• Development and Reproduction
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• v.10 no.3
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• pp.185-192
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• 2006

It is known that maternal diabetes induces a premature delivery and a congenital malformation. Although it was suggested that hyperglycemia is a main cause, little has been known. The effects of subtotal pancreactomized animal as a diabetic model on the embryo and the offspring were investigated. 1) Although survival rate of offsprings from the 90% pancreactomized female was lower than that of control, body weight of the survived offsprings was similar to that of control until 6 weeks after birth. When the liver was investigated, abnormal tissue was observed in both female and male offsprings from the pancreactomized mother. The level of major urinary protein(MUP) that is synthesized in the liver and secrets into urine was decreased in both female and male, supporting the liver abnormality. 2) In the pancreactomized female, size of the Langhan's island was increased and the liver also was observed abnormal. However, blood glucose level of the pancreactomized mother was not different from that of control. The low body weight and survival rate of offsprings from the $inhibin-{\alpha}$ overexpressed mice was very similar to those of the offsprings from diabetic mother. Thus when the mother itself was investigated after DNA injection, blood glucose level in the $inhibin-{\alpha}$ overexpressed mice was not different from that of control although size of the Langhan's island was decreased. Taken together, subtotal pancreactomy affected the offspring and embryo and influenced deeply the mother itself. However glucose level of the mother was not changed during investigation. It suggested that the relationship between the diabetic mother and embryo through offspring is very complex.

• Journal of Genetic Medicine
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• v.6 no.1
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• pp.74-80
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• 2009

Purpose: To assess the value of first-trimester pregnancy-associated plasma protein-A (PAPP-A), nuchal translucency (NT) and second-trimester alpha-fetoprotein (AFP), human chorionic gonadotropin (hCG), unconjugated estriol (uE3), and inhibin-A in predicting pregnancy complications other than fetal aneuploidy. Materials and Methods: A retrospective study in 3,121 singleton pregnancies with integrated testing was performed at Kangnam CHA hospital between January 2005 and December 2006. Baseline characteristics, pregnancy outcomes, and serum marker levels were obtained by review of the medical records. We analyzed the data to identify associations between the integrated screening markers and adverse pregnancy outcomes. Statistical analyses were performed with the SPSS program. Results: In preterm labor and preeclampsia, high AFP, hCG, and inhibin-A levels and low PAPP-A and NT levels were found to be significantly correlated (P<0.05). Elevated second-trimester inhibin-A levels were associated with preeclampsia (odds ratio 2.843), low birth weight (odds ratio 1.446), and preterm labor (odds ratio 1.287), and while decreased first-trimester PAPP-A levels were associated with preeclampsia (odds ratio 0.51) and preterm labor (odds ratio 0.75). Conclusion: First- and second-trimester maternal serum markers screening can be used for predicting high-risk pregnancies.