• Anatomy and Cell Biology
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• v.57 no.1
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• pp.119-128
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• 2024

Glucocorticoids play a physiologic role in the adult male reproductive functions, modulating gonadal steroid synthesis and spermatogenesis, through the glucocorticoid receptor (GR). The expression of GR has been described in several key testicular cell types, including somatic cells and early germ cell populations. Nothing is known on GR in human spermatozoa. Herein, we explored the GR expression and its possible role in normal and testicular varicocele semen samples from volunteer donors. After semen parameter evaluation by macro- and microscopic analysis, samples were centrifuged; then spermatozoa and culture media were recovered for further investigations. By western blotting and immunofluorescence analyses we evidenced for the first time in spermatozoa the presence of GR-D3 isoform which was reduced in sperm from varicocele patients. By treating sperm with the synthetic glucocorticoid dexamethasone (DEXA), we found that survival, motility, capacitation, and acrosome reaction were increased in both healthy and varicocele samples. GR involvement in mediating DEXA effects, was confirmed by using the GR inhibitor mifepristone (M2F). Worthy, we also discovered that sperm secretes different cortisol amounts depending on its physio-pathological status, suggesting a defence mechanism to escape the immune system attach in the female genital tract thus maintaining the immune-privilege as in the testis. Collectively, our data suggests a role for glucocorticoids in determining semen quality and function, as well as in participating on sperm immune defensive mechanisms. The novelty of this study may be beneficial and needs to take into account in artificial insemination/drug discovery aimed to enhancing sperm quality.

• Journal of Animal Science and Technology
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• v.49 no.1
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• pp.15-24
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• 2007

The purpose of this study was to evaluate the presence and differential expression of connexin isoforms in the efferent ductules (ED) of male rat reproductive tract during postnatal development. Total RNA was isolated from the ED collected from rats at 1 week, 2 weeks, 1 month, 3 months, and 6 months of ages. Expression of six connexin mRNAs of 14 isoforms tested was detected by semi-quantitative RT-PCR. Fluctuation of mRNA levels of connexins 26 and 30 was found according to ages. A significant decrease of connexin 31.1 mRNA level was observed in the ED at 1 month of age. The highest levels of connexin 37 and 45 mRNAs were detected in the ED of early developmental period, while the expression of connexin 43 was the lowest at 1 week of age. The present study demonstrates differential regulation on expression of connexin isoforms in the rat ED in age-dependent manner.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.12
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• pp.1566-1577
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• 2010

Nandrolone, 19-nortestosterone, is a synthetic androgenic-anabolic steroid promoting muscle growth. Nandrolone is also present in pig meat and sera at non-negligible levels. A number of scientific reports have suggested a positive relationship between incidence of infertility and increased meat consumption in humans. The present study was designed to determine out the effect of feeding nandrolone on the testis of the male reproductive tract. Mixtures of food and nandrolone at different concentrations (0.005 ppm and 0.5 ppm) were supplied to pubertal male rats for 6 weeks. Body weight was recorded every week during the entire experimental period. At the end of the treatment, the testis, epididymis, and epididymal fat were collected and weighted. Sperm numbers in the caudal epididymis were counted. Differential gene or protein expression of steroidogenic enzymes in the testes among experimental groups was determined by semi-quantitative real-time PCR or western blotting analysis, respectively. Histological changes of the testis induced by nandrolone treatment were examined by hematoxylin and eosin staining. Immunohistochemical analysis was employed to detect changes in the localization of steroidogenic enzymes in the testes among experimental animals. There were no significant changes on body, testis, epididymis, and epididymal fat weights among experimental groups. A significant increase of caudal sperm number was found in the 0.5 ppm nandrolone-treated group. Histological examination of the testes noted a high frequency of germ cell sloughing in seminiferous tubules of 0.5 ppm nandrolone-treated rats. Even though transcript levels of $3{\beta}$-hydroxysteroid dehydrogenase (HSD) I, $17{\beta}$-HSD4, and $17{\alpha}$-hydroxylase were influenced by nandrolone treatments, protein levels of all molecules examined in the present study were not significantly affected. Immunohistochemical analysis showed no visible changes in the localization of steroidogenic enzymes in the testes among experimental groups. The current study showed that oral intake of nandrolone in male rats for 6 weeks did not cause significant damage to the testis. It is considered that a feeding effect of nandrolone on male fertility would not be remarkable.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.311-315
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• 2005

This study was conducted to investigate the effects of disregarding of heat detection on embryo production in superovulation of Hanwoo cows. Donors which showed 1 or 2 times of normal heat and had no abnormality of reproductive tract were selected The superovulation was performed injection of 2.5 mL FSH (Antorin R-10, Japan) 2 times on 5 days before next heat and continuously with the reduction of dose to 0.5 mL of first injection 2 times in a day for next 3 days. Otherwise, the donors of CIDR group were inserted CIDR plus (with the capsule of estradiol benzoate 10 mg) on Day 10 from standing heat for 9 days. On 6 days from insertion of CIDR, FSH was injected above same manners. The response according to the natural heat and CIDR were $82.2\%,\;89.7\%$, respectively. There were no difference between both treatments. The recovery rates of embryos were 7.7, 10.5, respectively and transferable embryos were 3.4, 3.8. There showed significant difference between both treatments (p<0.05). These results suggested that disregarding of heat detection in superovulation could be produced transferable embryos for embryo transfer and preserve the donors from the excess hormonal administration and maintain the economical lift span of genetically available Hanwoo donors.

• Applied Microscopy
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• v.29 no.2
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• pp.241-253
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• 1999

Fertilization pattern of north temperate bats is known to be unique for their sperm storage in the female reproductive tract during hibernation (e.g. Korean greater horseshoe bats). They copulate in fall but their ejaculated spermatozoa survive until the next spring. In another words they can persist to survive during long hibernation under the cold condition $(8\sim13^{\circ}C)$ and are to be fertilized with the ovum ovulated in the next spring, so called delayed fertilization. The present study was designed to observe morphological and functional changes of spermatozoa plasma membrane of the bats, hamsters which are hibernators, and mice which are non-hibernators in the room and the cold (bat-hibernation) temperatures and to confirm influence of the temperature on spermatozoa; survival rate, acrosome reaction rate, protein distribution, $Na^+-K^+-ATPase$ activities and scanning electron microscopic histochemistry. Based on the experimental results obtained in the present study, there were no significant morphological and functional differences in the spermatozoa plasma membrane in both the room and cold (bat-hibernation) temperatures and such an absence of difference suggests that the spermatozoa plasma membrane might play a pertinent role as a protector for consistent fertilization during and after hibernation.

• Journal of Embryo Transfer
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• v.22 no.4
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• pp.245-249
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• 2007

The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), $10{\mu}g/ml$ follicle stimulating hormone (FSH), $35{\mu}g/ml$ luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$ and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within $39.1{\sim}70%$ in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.

• Journal of the Korea Organic Resources Recycling Association
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• v.29 no.3
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• pp.27-33
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• 2021

CO I gene sequence analysis was applied to earthworms that had been used as test animals in toxicity test in Institute of Kyeongbook Agrochemicals and earthworms used as vermicomposting agents in the farm of Youngdong province to identify their species names. In terms of molecular species, the former was identified as Eisenia fetida and the latter was Eisenia andrei. Cocoons produced from Eisenia fetida was more than those from Eisenia andrei. And No. of adults developed from eggs of Eisenia fetida was more or less higher than those developed from eggs of Eisenia andrei. These results were contradictory to previous reports on two Eisenia spp.. When Eisenia fetida was crossed with Eisenia andrei, hybridized eggs were produced and adults were developed from those eggs, but cocoons and adults were much less than those from non-crossed Eisenia fetida or Eisenia andrei. This indicated that two Eisenia spp. were not distinctly different biological species because there was no complete 'reproductive isolation' between Eisenia fetida and Eisenia andrei. However, this also meant that Eisenia fetida and Eisenia andrei had already been on the tract of speciation.

• Journal of Veterinary Clinics
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• v.32 no.2
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• pp.168-173
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• 2015

This study estimated the degree of uterine inflammation and resumption of ovarian cyclicity in postpartum dairy cows. In addition, the prevalence of health problems during calving and the postpartum period was investigated. A total of 224 Holstein dairy cows from four dairy farms in Chungcheong Province were used for the study. Uterine discharge was scored on a 0 to 5 rank scale (metricheck score) using a metricheck instrument from 1 to 8 weeks postpartum and uterine cytology (neutrophil level) was examined at 4, 6 and 8 weeks postpartum to evaluate uterine inflammation. Resumption of ovarian cyclicity was evaluated based on progesterone analysis (${\geq}1ng/mL$) at 4, 6 and 8 weeks postpartum. Postpartum disorders were diagnosed by veterinarians in the research team. The metricheck score decreased linearly (p < 0.0001) from weeks 1 ($3.4{\pm}0.1$) to 4 ($2.0{\pm}0.1$) postpartum, and then remained at that level through week 8 postpartum ($1.6{\pm}0.1$, p > 0.05). The neutrophil levels determined by uterine cytology assays also decreased (p < 0.0001) from week 4 ($22.3{\pm}1.8%$) to 6 ($12.1{\pm}1.4%$) postpartum, and then remained at that level through week 8 postpartum ($9.2{\pm}1.4%$, p > 0.05). The proportion of cows exhibiting the ovarian cyclicity increased linearly (p < 0.0001) from weeks 2 (12.1%) to 8 (74.3%) postpartum. The prevalence of dystocia and retained placenta were 20.5% and 30.4%, respectively. The prevalence of ketosis, milk fever, abomasal displacement, septicemic metritis, endometritis, subclinical endometritis, pyometra, and ovarian cysts were 18.8%, 3.6%, 4.0%, 36.2%, 29.5%, 17.0%, 4.5%, and 14.7%, respectively. The prevalence of digestive problems and mastitis were 5.4% and 9.6%, respectively. The prevalence of culling and death were 4.0% and 3.6%, respectively. The prevalence of dystocia, retained placenta, septicemic metritis, endometritis, ovarian cysts, digestive problems, mastitis, and culling differed among farms (p < 0.05 to 0.01). In conclusion, proper examinations for uterine inflammation and ovarian cyclicity during 4 to 8 weeks postpartum are useful clinical tools to maintain herd reproductive health. Dystocia, retained placenta, septicemic metritis, and endometritis were predominant disorders in dairy cows of the survey area.

• Development and Reproduction
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• v.10 no.2
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• pp.105-113
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• 2006

Reactive oxygen species(ROS) generated in cellular metabolism have an effect on cell maturation and development. In human reproductive tract, oxidative injury by ROS may induce female infertility. Also, oxidative injury may be responsible for developmental retardation and arrest of mammalian preimplantation embryos. Activating transcription factor 4(ATF4) is a member of the cyclic-AMP response element-binding(CREB) familiy of basic region- leucine zipper(bZip). ATF4 is known to regulate stress response to protect cell from various stress factors and inducer of apoptisis. The purpose of this study was to investigate whether ATF4 is involved in the defensive mechanism in oxidative stress condition during the development of mouse preimplantation embryos. To verify the expression of ATF4 in oxidative stress condition, 2-cell stage embryos were cultured in HTF media containing 0.1mM, 0.5mM or 1mM hydrogen peroxide($H_2O_2$) for 1hr(2-cell), 8hr(4-cell), 17hr(8-cell), 24hr(morula), 48hr(early blastocyst) or 64hr(late blastocyst). The developmental rate decreased in the 0.1mM $H_2O_2$ treated group compared with control group. In embryos treated with 0.5mM and 1mM $H_2O_2$ showed 2-cell block. As a results of the semi-quantitative RT-PCR analysis of SOD1, ATF4 and Bax gene expression, SOD1, ATF4 and Bax genes were increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. In 2-cell embryos, expression of SOD1, ATF4 and Bax genes were notably increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. Immunofluorescence analysis showed that ATF4 protein was localized at the cytoplasm of preimplantation embryos. The increase in ATF4 immunoreactivety was observed in the 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. It suggests that oxidative stress by $H_2O_2$ induces expression of ATF4 and may be involved in protection mechanism in preimplantation embryos from oxidative injury.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.63-67
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• 2007

This study was performed to evaluate the efficacy of sodium carboxymethylcellulose (SCMC) for the prevention of postoperative uterus and ovary adhesion formation in the Korean black goats. Twenty adult female Korean black goats randomly were divided into five groups with four animals in each group. After the routine laparotomy, catheter for injection was inserted into the abdomen. Before abdominal closure, saline, 1% SCMC, 2% SCMC or 0.4% HA solution (50ml/10kg of body weight/head) were injected in the abdominal cavity in each group. Three weeks after surgery, second laparotomy was performed and the adhesions were scored on a scale of 0 to 10 according to their vascularity and adhesion size in uterus and ovary. This trial was repeated the four times with the interval of three weeks. In the first and second trial, the group treated with 2% SCMC significantly reduced the adhesion formation than other treatment groups (P<0.05). In the third trial, the adhesion formation was significantly reduced in 2% SCMC and 1% SCMC (P<0.05). In the fourth trial, 2% SCMC reduced the adhesion formation. However, there was no significant difference among other groups. This study showed that the 2% SCMC administered at the end of the surgery reduced the adhesion formation in the Korean black goats.