• Journal of Animal Science and Technology
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• 제51권6호
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• pp.485-492
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• 2009

Estrogen has dramatic effects on the development and function of the reproductive tract in mammals. Although diethylstilbestrol (DES) triggers the development of reproductive organs in immature animals, continued exposure to DES induces dysfunction of the female reproductive tract in mice. To investigate the effects of neonatal estrogen exposure on the reproductive tract of female chickens, we implanted DES pellets into the abdominal region of immature female chicks and then examined the effects of DES on the oviducts of both immature chicks and sexually mature chickens (30 weeks old). DES induced mass growth and differentiation of the oviduct in immature chicks. The chick oviduct increased by 2.7- and 29-fold in length and weight, respectively, following primary DES stimulation. In secondary DES stimulation, the length and weight of the chick oviduct increased by 4.5- and 74-fold, respectively. Additionally, DES treatments caused abnormal development of the infundibulum and magnum in hen oviducts. Furthermore, infundibulum abnormality gave rise to unusual ovulation of follicles and resulted in infertility and dysfunction of the magnum, such as less production of egg white proteins. Our results indicate that DES exposure during early developmental stages in chickens has detrimental effects on the development and maintenance of the female reproductive tract after sexual maturation.

• Journal of Animal Science and Technology
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• 제63권4호
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• pp.766-777
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• 2021

Bezafibrate, a fibrate drug used as a lipid-lowering agent to treat hyperlipidemia, is a pan-agonist of peroxisome proliferator-activated receptor alpha. It can enhance mitochondrial fatty acid oxidation, oxidative phosphorylation, and mitochondrial biogenesis. After ovulation, oocytes may get arrested at the metaphase II (MII) stage until fertilization beyond optimal timing, which is termed as post-ovulatory aging. Post-ovulatory aging is a disease that degrades DNA, mitochondria, and oxidative system, and has a negative impact on embryo development and quality; however, the impact of bezafibrate during post-ovulatory aging has not been fully defined. In the present study, we assessed the ability of bezafibrate to prevent the progression of aging in in vitro conditions as well as the underlying mechanisms in pigs. An appropriate concentration of this drug (50 µM) was added, and then oxidative stress, reactive oxygen species downstream, mitochondrial biogenesis, and mitochondrial function were analyzed via immunofluorescence staining and real-time polymerase chain reaction. Bezafibrate significantly alleviated reactive oxygen species and ameliorated glutathione production simultaneously in oocytes and embryos. Moreover, it diminished H2A.X and attenuated CASPASE 3 expression produced by oxidative stress in oocytes and embryos. Furthermore, bezafibrate remarkably improved the mitochondrial function and blastocyst quality as well as markedly reduced the mitochondria/TOM20 ratio and mtDNA copy number. The elevated PARKIN level indicated that mitophagy was induced by bezafibrate treatment after post-ovulatory aging. Collectively, these results suggest that bezafibrate beneficially affects against porcine post-ovulatory oocyte aging in porcine by its antioxidant property and mitochondrial protection.

• Anatomy and Cell Biology
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• 제56권4호
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• pp.526-537
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• 2023

Hepatitis C virus (HCV) infection is a major health problem worldwide and its eradication is mandatory. Direct acting HCV polymerase inhibitors, such as Sofosbuvir (SOF), is an effective regimen. However, it has some side effects like mutagenesis, carcinogenesis, and the impairment of testicular function. It is important to evaluate the safety of SOF on the ovary, as there are no studies yet. Increasing the production of Reactive Oxygen Species (ROS), causes oxidative stress, which affects ovulation process, female reproduction, and fertility. Accumulation of SOF in the cells was demonstrated to promote ROS generation. Vitamin E (Vit E) is an antioxidant agent that has an essential role in the female reproductive system, its deficiency can cause infertility. We explored the effect of SOF treatment alone and co-treated with Vit E on ovarian ROS level and ovarian morphology experimentally using biochemical and immunohistochemical studies. Significant changes in oxidative stress markers; nitric oxide and malondialdehyde lipid peroxidation, antioxidant enzymes; catalase, super oxide dismutase, and reduced glutathione, proliferating markers; proliferation cell nuclear antigen and Ki-67 antigen and caspase 3 apoptotic marker were demonstrated. It was shown that where SOF induced oxidative stress, it also aggravated ovarian dysfunction. The essential role of Vit E as an antioxidant agent in protecting the ovarian tissue from the effect of oxidative stress markers and preserving its function was also displayed. This could be guidance to add Vit E supplements to SOF regimens to limit its injurious effect on ovarian function.

• Clinical and Experimental Reproductive Medicine
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• 제32권2호
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• pp.155-164
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• 2005

Objective: This study was performed to investigate the effects of pioglitazone, an insulin sensitizing agent, on insulin resistance, ovarian function and intraovarian stromal blood flow in patients with polycystic ovarian syndrome (PCOS). Material and Methods: Thirty patients with PCOS, aged 18~34 years, were recruited. Criteria for diagnosis of PCOS were as defined in 2003 Rotterdam consensus. They were treated for 6 months with pioglitazone at a dose of 30 mg/day orally. The hormonal blood profile, fasting serum glucose levels, a glycemic response to 75 g oral glucose tolerance test (OGTT), and an ovarian stromal artery (OSA) blood flow were assessed at baseline and after 6 months of treatment. Results: Eighteen (60.0%) of 30 patients treated with pioglitazone demonstrated a spontaneous ovulation After pioglitazone treatment, fasting insulin concentrations, serum glucose levels after 75 g OGTT significantly decreased (p=0.001, p=0.04, respectively), and fasting glucose to insulin (G/I) ratio significantly increased (p<0.001). The pioglitazone treatment induced a significant reduction in serum LH, testosterone (T) and free T levels (p<0.001, p=0.02, p=0.002, respectively). The resistance index (RI) values of OSA significantly increased after treatment (p<0.001). In analyzing pioglitazone-treated patients according to their body mass index (BMI), nonobese group as well as obese group showed a significant improvement in fasting G/I ratio (p<0.01). The pioglitazone treatment induced a significant reduction in serum LH and free T levels in nonobese group (p<0.001, p<0.05, respectively) as well as obese group (p=0.001, p<0.05, respectively). The RI values of OSA significantly increased in both nonobese and obese groups after pioglitazone treatment (p<0.001, p=0.003, respectively). Conclusions: Pioglitazone could ameliorate the glycoinsulinemic metabolism, and this beneficial effects of this drug could improve the endocrine-reproductive condition associated with the decrease of ovarian stromal artery blood flow, in both nonobese and obese patients with PCOS.

• 한국수정란이식학회지
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• 제22권1호
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• pp.53-61
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• 2007

본 연구는 PI한 한국흑염소 수정란의 이식 결과를 통해 수란 흑염소의 수태율에 영향을 미칠 것으로 생각되는 여러 가지 요인을 분석함으로써, 높은 수태율을 얻을 수 있는 수란 흑염소의 최적 조건을 찾아낼 목적으로 수행하였다. 분석 결과, 수태율에 유의적인 영향을 주는 요인들은 발정형태, 수술 빈도, 이식 부위, 황체의 발육 단계, 수정란의 발육단계, 이식된 수정란의 수 등이었다. 자연 발정이 관찰되어 이식된 흑염소(59.1%, 13/22)들이 $CIDR^(R)$로 발정이 유도된 후 이식된 흑염소(36.8%, 118/321)에서보다 높은 수태율을 나타내었으며(P<0.05), 두 번째 수술 받은 흑염소의 수태율(56.5%, 13/23)이 처음 이식 받은 흑염소(36.5%, 116/318)에 비해 수태율이 높았다(P<0.05). 이식한 부위에 따른 수태율은 좌측 난관에 이식한 흑염소(49.0%, 50/102)가 오른쪽 난관에 이식한 흑염소(35.9%, 46/128)에 비해 수태율이 더 높게 나타났으며(P<0.05), 황체의 발육 단계에 따른 수태율에서는 $CH_1$단계(47.5%, 57/120) 출혈체를 가진 수란 흑염소에서 $CH_3(17.9%,\;7/39)$의 출혈체를 가진 수란 흑염소보다 높은 수태율을 얻었다(P<0.01). 수정란의 발육단계에 따른 차이는 난관 이식의 경우에 1세포기 배가 이식된 수란 흑염소의 수태율(51.6%, 49/95)이 4세포기배를 이식한 경우(24.5%, 12/49)보다 높았다(P<0.01). 수정란의 수는 2개를 이식했을 때(27.%, 37/137)보다 3개를 이식했을 때(47.6%, 50/105) 더 높은 수태율을 얻었다(P<0.01). 수태율에 유의적인 영향이 없는 요인들은 난관 유착이나 자궁 유착, 난소 유착, 자궁각의 크기, 황체의 수, 대형 난포의 존재 유무, 이식의 난이도 등이었다 그러나, 중간 크기의 자궁을 가진 수란 흑염소(38.9%, 122/314)에서 직경 5mm 이하의 작은 자궁을 가진 수란 흑염소(20%, 1/5)나 20mm 이상의 큰 자궁을 가진 수란 흑염소(18.2%, 2/11)보다 수태율이 높은 경향을 보였고, 배란 황체가 있는 같은 쪽 난소에 대형 난포가 존재할 경우(53.3%, 16/30)에 존재하지 않는 경우(37.1%, 104/280)보다 수태율이 높아지는 경향을 보였으며, 이식이 쉽게 이루어진 경우(39.2%, 125/319)에 이식이 어렵거나(27.8%, 5/18) 곤란한 경우(0%, 0/3)에서보다 높은 수태율을 얻을 수 있었다. 따라서, 자궁각의 크기나 대형 난포의 존재 유무, 이식의 난이도 등도 수정란 이식 후의 수태율에 영향을 줄 수 있을 것으로 생각된다. 이상의 결과로 볼 때 PI한 한국 흑염소 수정란의 이식 시 높은 수태율을 얻기 위해서는 수란 흑염소는 자연 발정 온 개체를 이용하고, 난관에 이식하고자하는 경우에는 난소에 $CH_1$ 단계의 출혈체가 존재하는 쪽 난관에 1세포기의 수정란을, 그리고 자궁에 이식하는 경우에는 난소에 발육 단계가 $CL_3$인 황체가 존재하는 쪽 자궁각에 중기 배반포나 후기 배반포를 이식하는 것이 가장 바람직하다는 결론을 얻었다.

• 한국수정란이식학회지
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• 제24권3호
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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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• 제31권1호
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• pp.59-65
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• 2004

Objectives: The aim of this study was to assess toxicities of cryoprotectants. Methods: Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and 1, 2-propanediol (PROH), were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or PROH. Embryo development was evaluated up to the blastocyst stage. Blastocysts were stained with bis-benzimide to evaluate the cell count and with terminal deoxynucleotidyl transferase mediated dUTP nick labeling (TUNEL) to assess apoptosis. Results: The total cell count of blastocysts that were treated with DMSO at the 2-cell stage was significantly lower than that were treated with PROH ($75.9{\pm}27.0$) or the control ($99.0{\pm}18.3$) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the PROH treated group (p<0.05). Both DMSO ($14.2{\pm}1.5$) and PROH ($11.2{\pm}1.4$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.2{\pm}0.9$, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the PROH treated group (p<0.001). Conclusions: The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or PROH at room temperature. When comparing two cryoprotective agents, PROH appeared to be less toxic than DMSO at least in a murine embryo model.

• 대한수의학회지
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• 제29권3호
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• pp.373-381
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• 1989

The aim of the present study was to test the efficiency of estrous induction in the premature, metestrous and anestrous bitches. The estrus was induced with prostaglandin $F_{2{\alpha}}$, estradiol-$17{\beta}$, pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin(HCG) in the treatment A, and with PMSG and HCG in the treatment B. Day 0 was the first day of estrone injection in the treatment A and the day of PMSG injection in the treatment B. Twenty three of the twenty six bitches were laparotomized under general anesthesia between 11 and 18 days after onset of behavioral estrus, whereas three bitches were not laparotomized and remained until parturition. Ovarian responses were evaluated with the total number of corpora lutea or ovulation sites. The uterine horns were flushed with phosphate-buffered saline added heat treated canine serum(10%), the flushing media was collected into watch glass and the ova were examined under stereomicroscope. The results obtained were as follows: 1. Standing estrus was observed on the day $17.7{\pm}1.5$ after injection of estrone in the treament A, but ovarian responses were not detectable. 2. Standing estrus was observed on the day $12.2{\pm}0.2$ after injection of PMSG in the treament Band 14 of 17 bitches showed ovarian responses. Ova were recovered in 9 of the 14 bitches. 3. Ovarian responses were observed in one of the three premature bitches. two of the three metestrous bitches and all of the 11 anestrous bitches. The average number of the ova collected from 9 bitchs were $12.2{\pm}1.4$. 4. Three bitches in the treament B exhibited behavioural estrus and all of them were mated with fertile male dog, resulting the pregnancy in only one bitch. The pregnant bitch gave the birth of two pups.

• 한국가축번식학회지
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• 제11권3호
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• pp.155-160
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• 1987

This study was carried out to determine the effects of cryoprotectants, freezing and thawing rates on the survival of 8-cell mouse embryos. The female ICR mice were induced to superovulated by intraperitoneal injections of 5 i.u. PMSG and 5 i.u. HCG given 48h apart and then were paired with males of the same strain. They were killed and embryos were flushed from the oviducts and uteri on 3 days after injection of HCG. Embryos were flushed with modified Dulbecco's phosphate buffered saline and equilibrated with 1.5M-dimethyl sulfoxide (DMSO) or 1.5M-glycerol by 3-step procedure. The freezing rates of the embryos were 1$^{\circ}C$/min from room temperature to -5$^{\circ}C$ and the embryos were seeded at -5$^{\circ}C$. After being held for 3 min at the seeding temperature, the rates were 0.3$^{\circ}C$/min from -5$^{\circ}C$ to -35$^{\circ}C$. From -35$^{\circ}C$ to -7$0^{\circ}C$, the rates were divided into 0.1$^{\circ}C$/min, 1$^{\circ}C$/min and 1$0^{\circ}C$/min, respectively. After being held for 5 min at -7$0^{\circ}C$, the embryos were plunged directly into liquid nitrogen. The embryos were thawed at 4$^{\circ}C$/min and 12$^{\circ}C$/min from -196$^{\circ}C$ to 37$^{\circ}C$, and for 2 min in 37$^{\circ}C$ water bath, respectively. The average number of ovulation points and embryos recovered were 42.7 and 34 appearing 79.5% recovery rate. Eight cell embryos in the embryos recovered were 26.3. The survival rates of embryos according to the freezing rates in the presence of 1.5M-DMSO were 73.5~80.6% at 0.1$^{\circ}C$/min, 75.0~79.5% at 1$^{\circ}C$/min and 52.8~54.7% at 1$0^{\circ}C$/min, but in the presence of 1.5M-glycerol were 62.9~67.6% at 0.1$^{\circ}C$/min, 61.4~68.3% at 1$^{\circ}C$/min and 25.5~30.2% at 1$0^{\circ}C$/min. The survival rates of embryos were not affected by the thawing rates.

• Reproductive and Developmental Biology
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• 제39권3호
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• pp.77-81
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• 2015

본 연구에서 배아의 생식세포 동결에 가장 흔히 쓰이고 있는 두 가지 동결 보호제, 즉 DMSO와 EG의 독성을 비교하고자 생쥐 수정란 모델을 이용한 실험을 하였다. 생후 6주령의 암컷 생쥐 $F_1$ hybrid mice에 10 IU의 PMSG를 복강 주사하여 과배란을 유도하고, 2-세포기 배아를 획득하고 DMSO와 EG 각각 노출시킨 후, 배양을 하였다. 배반포의 전체 세포수는 2-세포기 단계에서 DMSO($68.1{\pm}24.1$)로 EG($81.2{\pm}27.0$) 혹은 control($99.0{\pm}18.3$)(p<0.001) 처리구에 비해서 유의적으로 낮았다. DMSO 처리구가 EG 처리구에 비해 세포수가 적었다. DMSO($15.4{\pm}1.5$)와 EG($10.2{\pm}1.4$) 두 처리구는 대조구($6.1{\pm}0.9$, p<0.0001)와 비교해서 배반포에서 세포사 비율이 더 높음을 확인했다. 또한, DMSO 처리구는 EG 처리구(p<0.001)보다 더 많은 세포사멸된 세포가 확인되었다. DMSO 또는 EG 처리군과 대조군 사이에는 배아 부화율에 있어서 차이가 있었으며, 이는 배아에 대한 동결 보호제의 잠재적인 독성을 확인한 결과였다. 이번 연구에서 장기간 처리했을 때 EG 처리군보다 DMSO 처리군에서 배아발달과 세포수가 저하된 것은 DMSO의 독성이 더 높을 것으로 사료된다.