• Laboratory Medicine Online
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• v.7 no.2
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• pp.83-87
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• 2017

Pseudohypoparathyroidism (PHP) is a rare disorder caused by genetic and epigenetic aberrations in the GNAS complex locus resulting in impaired expression of stimulatory G protein ($Gs{\alpha}$). PHP type Ib (PHP-Ib) is characterized by hypocalcemia and hyperphosphatemia due to renal resistance to the parathyroid hormone, and is distinguished from PHP-Ia by the absence of osteodystrophic features. An 11-yr-old boy presented with poor oral intake and cramping lower limb pain after physical activity. Laboratory studies revealed hypocalcemia, hyperphosphatemia, and increased parathyroid hormone levels. The GNAS complex locus was evaluated using the methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay. Gain of methylation in the NESP55 domain and loss of methylation in the antisense (AS) transcript, XL, and A/B domains in the maternal allele were observed. Consequently, we present a case of PHP-Ib diagnosed using MS-MLPA.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.2
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• pp.101-111
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• 2005

배 경: Pituitary adenylate cyclase-activating polypeptide (PACAP)은 양의 시상하부에서 추출된 신경펩타이드 호르몬으로 난소에도 존재하여 배양된 과립막 세포에서 스테로이드합성과 cyclic AMP 형성을 촉진함이 보고되었다. 목 적: 흰쥐 난소를 실험 모델로 사용하여 배란시 황체화호르몬 (luteinizing hormone; LH)에 의해 유도된 PACAP과 PACAP 수용체의 유전자 발현양상과 신호 전달경로를 규명하고자 하였다. 재료 및 방법: 미성숙 흰쥐의 배란전 난포를 체외 배양하면서 LH로 처리하고 PACAP 및 PACAP수용체의 유전자 발현을 보기 위해서는 Northern blot 분석과 in situ hybridization (ISH)을, 그리고 단백질 수준의 PACAP 검색을 위해서는 enzyme linked immunosorbent assay (ELISA) 분석을 이용하였다. 결 과: LH 처리 후 Northern blot상의 PACAP 유전자 발현은 6~9시간에 일시적으로 최고치에 도달하였으며 ISH로 보아 과립막 세포에서 발현됨을 알 수 있었다. ELISA 분석 상 PACAP 단백질도 LH처리 후 6~12시간에 최고치를 나타내었으며, PACAP 수용체 mRNA 역시 3~9시간에 최고치로 과립막 세포에서 발현되었다. Adenylate cyclase (AC) 억제제인 MDL12330A 처리시 LH로 발현된 PACAP mRNA가 감소되며, AC의 활성제인 forskolin 처리에는 LH시와 유사한 PACAP mRNA의 발현양상을 나타내었다. 그러나 protein kinase C (PKC)의 억제제인 chelerythrine과 2-0-tetradecanolphorbol-13-acetate (TPA) 처리로는 PACAP 의 유전자 발현에 영향을 주지 못하였다. 5-lipoxygenase의 억제제인 MK886이나 nordihydroguaiaretic acid (NDGA)로 처리한 결과 LH로 유도된 PACAP 유전자의 발현이 감소되었으나, cyclooxygenase의 억제제인 indomethacin은 별로 영향을 주지 못하였다. MEK와 p38의 억제제인 PD98059와 SB203580도 LH로 촉진 된 PACAP의 유전자 발현을 농도 의존적으로 억제하였다. 결 론 : 배란전 난포에서 PACAP과 PACAP 수용체의 유전자 발현은 모두 LH의 폭발적 분비에 의해 유도되어 일시적으로 과립막 세포에서 나타나 배란을 위한 국소적인 조절 작용을 할 것으로 추정되며, LH로 촉진된 PACAP 유전자 발현을 위한 신호전달은 cAMP-PKA, lipoxygenase 및 MAP kinase 경로를 통하는 것으로 사료된다.

• YAKHAK HOEJI
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• v.44 no.5
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• pp.391-398
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• 2000

Leptin, the product of the ob gene, is a small peptide molecule synthesized by white adipocytes with an important role in the regulation of body fat and food intake. Based on the evidence that synthesis of leptin is regulated by female sex hormone, estrogen, this present study was investigated whether sex hormone precursor DHEA, can regulate obese gene expression in lean and genetically obese (ob/ob) mice. Antiobesity activity of DHEA was evaluated by determining body weight, food consumption, epididymal fat weight and serum levels of cholesterol and triglyceride in ICR, C57BL/6J, and ob/ob mice. The treatment of C57BL/6J lean and obese mice with a diet containing 0.3% and 0.6% DHEA resulted in lowered rates of weight gain in comparison to non-treated mice, although much greater response was found in the obese mice. All other concentrations of DHEA (0.015%, 0.06%, 0.15%, 0.3%) except the highest one(0.6%) showed no significant effects on weight gain in ICR mice. Food consumption was significantly decreased in all mice treated with 0.6% DHEA, whereas it was not decreased in ICR mice at lower concentrations than 0.6% DHEA. DHEA decreased significantly epididymal adipose tissue weight and serum triglyceride levels dose dependently in lean and obese mice. However serum cholesterol levels were decreased at lower concentrations than 0.15% DHEA and increased at concentrations of 0.3% and 0.6% DHEA in lean and obese mice. These increases in serum cholestrol levels at high concentrations of DHEA might result from the fact that DHEA has a cholesterol moiety thereby interfered the assay system. As an approach to elucidate the mechanism for antiobesity activity of DHEA, we examined mRNA levels of obese gene in the adipocyte and obese gene product (leptin) in the serum. The results showed that DHEA did not affect obese gene expression in ICR and C57BL/6J mice. Therefore, we concluded that antiobesity activity of DHEA was not modulated by obese gene expression.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.3
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• pp.245-254
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• 2021

Objective: In humans, polycystic ovary syndrome (PCOS) is an androgen-dependent ovarian disorder. Aberrant gene expression in folliculogenesis can arrest the transition of preantral to antral follicles, leading to PCOS. We explored the possible role of altered gene expression in preantral follicles of estradiol valerate (EV) induced polycystic ovaries (PCO) in a mouse model. Methods: Twenty female balb/c mice (8 weeks, 20.0±1.5 g) were grouped into control and PCO groups. PCO was induced by intramuscular EV injection. After 8 weeks, the animals were killed by cervical dislocation. Blood serum (for hormonal assessments using the enzyme-linked immunosorbent assay technique) was aspirated, and ovaries (the right ovary for histological examinations and the left for quantitative real-time polymerase) were dissected. Results: Compared to the control group, the PCO group showed significantly lower values for the mean body weight, number of preantral and antral follicles, serum levels of estradiol, luteinizing hormone, testosterone, and follicle-stimulating hormone, and gene expression of TGFB1, GDF9 and BMPR2 (p<0.05). Serum progesterone levels were significantly higher in the PCO animals than in the control group (p<0.05). No significant between-group differences (p>0.05) were found in BMP6 or BMP15 expression. Conclusion: In animals with EV-induced PCO, the preantral follicles did not develop into antral follicles. In this mouse model, the gene expression of TGFB1, GDF9, and BMPR2 was lower in preantral follicles, which is probably related to the pathologic conditions of PCO. Hypoandrogenism was also detected in this EV-induced murine PCO model.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.426-439
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• 2021

It is already well known that castration improves marbling quality but exact timing of castration is still highly debated in beef cattle production industry. After castration, blood hormonal changes occur in steer and objective of this study was to investigate the effects of growth hormone (GH) levels on adipocyte differentiation in stromal vascular cells (SVCs) and transdifferentiation into adipocytes in C2C12 myoblasts. Total GH concentrations were measured via enzyme-linked immunosorbent assay (ELISA) in 24 male calves and 4 female calves. Cell proliferation, cellular triglyceride (TG) accumulation, and the cell's lipolytic capability were measured in C2C12 myoblasts and SVCs. Myogenic, adipogenic, and brown adipocyte-specific gene expression was measured via real-time polymerase chain reaction (PCR) using SYBR green. Serum GH levels were the highest in late-castrated calves. Treatment with 5 ng/mL GH resulted in greater TG accumulation as well as increased CCAAT-enhancer-binding protein (C/EBP)α and peroxisome proliferator-activated receptor (PPAR)γ expression compared to that after treatment with 15 ng/mL GH. Treatment with 5 ng/mL GH also resulted in lower myogenin (myo)G and myoD expression compared to that after treatment with 15 ng/mL GH. The expression of bone morphogenetic protein (BMP) 7 after treatment with 5 ng/mL GH was higher than that after treatment with 15 ng/mL GH. But carcass characteristics data showed no significant difference between early and late castrated steers. Therefore, our results indicate that castration timing does not seem to be inevitable determinate of carcass qualities, particularly carcass weight and marbling score in Hanwoo beef cattle.

• The Korean Journal of Physiology and Pharmacology
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• v.27 no.2
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• pp.167-176
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• 2023

This study aims to explore the impact of Rehmannioside D (RD) on ovarian functions of rats with diminished ovarian reserve (DOR) and its underlying mechanisms of action. A single injection of cyclophosphamide was performed to establish a DOR rat model, and fourteen days after the injection, the rats were intragastrically administrated with RD for two weeks. Rat estrus cycles were tested using vaginal smears. Ovarian tissues were histologically evaluated, the number of primordial, mature, and atretic follicles was calculated, and the apoptotic rate of granulosa cells. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E₂) levels were determined by ELISA assays. Protein levels of Forkhead Box O1 (FOXO1), KLOTHO, Bcl-2, and Bax were investigated in ovarian tissues of DOR rats. The binding between FOXO1 and KLOTHO was verified by ChIP assay. High-dose administration of RD into DOR rats improved their estrus cycles, increased ovarian index, enhanced the number of primordial and mature follicles, reduced the number of atretic follicle number, and ovarian granulosa cell apoptosis in addition to inhibiting FSH and LH levels and upregulating E₂ expression. FOXO1 and KLOTHO were significantly suppressed in DOR rats. FOXO1 knockdown partially suppressed the protective effects of RD on DOR rats, and KLOTHO overexpression could restore RD-induced blockade of DOR development despite knocking down FOXO1. FOXO1 antibody enriched KLOTHO promoter, and the binding between them was reduced in DOR group compared to that in sham group. RD improved ovarian functions in DOR rats and diminished granulosa cell apoptosis via the FOXO1/KLOTHO axis.

• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1351-1360
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• 2023

Endocrine-disrupting chemicals (EDCs) are compounds that disturb hormonal homeostasis by binding to receptors. EDCs are metabolized through hepatic enzymes, causing altered transcriptional activities of hormone receptors, and thus necessitating the exploration of the potential endocrine-disrupting activities of EDC-derived metabolites. Accordingly, we have developed an integrative workflow for evaluating the post-metabolic activity of potential hazardous compounds. The system facilitates the identification of metabolites that exert hormonal disruption through the integrative application of an MS/MS similarity network and predictive biotransformation based on known hepatic enzymatic reactions. As proof-of-concept, the transcriptional activities of 13 chemicals were evaluated by applying the in vitro metabolic module (S9 fraction). Identified among the tested chemicals were three thyroid hormone receptor (THR) agonistic compounds that showed increased transcriptional activities after phase I+II reactions (T3, 309.1 ± 17.3%; DITPA, 30.7 ± 1.8%; GC-1, 160.6 ± 8.6% to the corresponding parents). The metabolic profiles of these three compounds showed common biotransformation patterns, particularly in the phase II reactions (glucuronide conjugation, sulfation, GSH conjugation, and amino acid conjugation). Data-dependent exploration based on molecular network analysis of T3 profiles revealed that lipids and lipid-like molecules were the most enriched biotransformants. The subsequent subnetwork analysis proposed 14 additional features, including T4 in addition to 9 metabolized compounds that were annotated by prediction system based on possible hepatic enzymatic reaction. The other 10 THR agonistic negative compounds showed unique biotransformation patterns according to structural commonality, which corresponded to previous in vivo studies. Our evaluation system demonstrated highly predictive and accurate performance in determining the potential thyroid-disrupting activity of EDC-derived metabolites and for proposing novel biotransformants.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.169-176
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• 2009

Domestic bitches are non-seasonally monoestrus; spontaneously ovulate only once or twice occurs at anytime of the year. Estrus induction has been applied infrequent estrus, misleading ovulation, mating difficulties, failure to conceive after normal mating, pregnancy failure and biological research. Protocol of estrus induction which included variable hormones such as FSH, GnRH, and PMSG have been applied for the last decades. Recently, Bromocriptine, one of anti-prolatin/dopamine agonist has been occasionally applied for estrus induction. The study was carried out to investigate the effective method for the induction of estrus in bitches using different hormone treatments, and the initiation time of estrus from hormone treatment by assessments of cytological observation and blood plasma progesterone concentration. A total of 54 bitches on anestrus were selected for the study and divided randomly into 8 treatment groups as follow. Control, natural estrus; FSH (L), FSH (1.5 mg/kg, twice a day, $Falltrophin^{(R)}$, Vetrepharm); FSH (H), FSH (3.0 mg/kg, twice a day); GnRH+FSH, GnRH (5 ug/kg, once first day, $GNADON^{(R)}$, Dongbang)+FSH (3 mg/kg, SID); PMSG, PMSG (50 IU/kg, every third day $FOLLIGON^{(R)}$, Intevet); GnRH+PMSG, GnRH (5 ug/kg, only first day)+PMSG (50 IU/kg, every third day); GnRH, GnRH (5 ug/kg, only first day); Bromocriptine, bromocriptine (0.3 mg/kg, SID, $Parlodel^{(R)}$, Novartis). The bitches were evaluated clinical sign, cytological exam and $OVUCHECK^{(R)}$ Premate for assessment of estrus induction. Estrus induction rates were significantly (P<0.05) higher in GnRH+PMSG (100%) compared to others. PMSG and GnRH+PMGS (87.5 and 100%) and Bromocriptine (77.8%) were higher than others except GnRH+PMSG. Analysis of vaginal smear has proved to be effective a correct assessment of estrus induction with assay of progesterone concentration by $OVUCHECK^{(R)}$ Premate. Proestrus initiated by the $6^{th}$ after induction in most case. In conclusion, bromocriptine is an effective drug for estrus induction in bitches and assay of progesterone concentration by $OVUCHECK^{(R)}$ Premate with examination of vaginal smear that should be useful to detection of estrus induction of estrus induced bitches.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.2
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• pp.115-123
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• 2006

Objective: To investigate new signal transduction cascade through integrin, FAK and ERK in the suppressed cell proliferation by GnRH-I and -II. Method: Human endometrial cancer cells (HEC1A) were cultured under the following condition: DMEM/F12 (10% FBS). GnRH-I and -II were treated time (0, 5, 10, 15, 20, 30 min; 100 nM) and dose (10 nM or 100 nM; 20 min) dependent manner according to experimental purposes. Cell proliferation was measured using [$^3H$] thymidine incorporation assay. Immunoblotting was utilized to detect proteins. Results: GnRH-I and -II inhibited proliferation of HEC1A cells and induced expression of integrin ${\beta}3$. Phosphorylation of FAK and ERK were induced by GnRH-I and -II. Conclusion: GnRH inhibited cell proliferation via the expression of integrin and FAK, ERK phosphorylation.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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• pp.149-155
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• 1992

The present study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone(20$\alpha$-OHP) levels before and after parturition, 4 pluriparous goats were offered for this experiment. Blood samples were taken from jugular vein on Days, 5, 3, 2 and 1 before parturition, the day of parturition, 1, 3, 5, 7 and 9 after parturition. The blood samples were centraifuged and stored at -2$0^{\circ}C$ until hormone assay. The serum levels of progesterone and 20$\alpha$-OHP were measrued by radioimmunoassay. The changes of serum progesterone level during peripartum period were characterized as a remarkable decrease. The progesterone level was 4.05$\pm$0.52ng/ml on 56 days before parturition and decreased to 2.24$\pm$0.38ng/ml on 1 day before parturition and 0.79$\pm$0.09ng/ml on the day of parturition and the basal level was maintained through 9 days of postpartum period. The serum level of 20$\alpha$-OHP during the peripartum period was 1.25$\pm$0.21ng/ml on 5 days before paturition and increased to 1.32$\pm$0.25 on 3 days and 1.59$\pm$0.24ng/ml on 1 day before parturition, and reached a peak level of 1.78$\pm$0.25ng/ml just prior to parturition and then decreased greatly to 0.31$\pm$0.03ng/ml on 1 day postpartum and the basal level was remained until 9 days postpartum. The high serum level of 20$\alpha$-OHP, which was peak just prior to parturition, was maintained for 2 days following the onset of remarkable decrease in the serum level of progesterone. From the above results, it was concluded that the enzyme 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) catalyzing the conversion of progesterone to a biologically inactive steroid, 20$\alpha$-OHP was active properly in the luteal cells in Korean native goats.