• The Korean Journal of Zoology
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• v.32 no.3
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• pp.264-274
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• 1989

In order to study the influence of spermatozoa and testicular fluid on the component and composition of proteins in epididymal fluid of mice, histological differentiation of testis and epididymis were observed during sexual maturation, and the proteins in epididymal fluids collected according to the characteristics of lumination were analyzed by electrophoresis (SDS-PAGE). In 10 day-old mouse, both of,testis and epididymis were undifferentiated. In 20 day-old mouse, epididymis was primitively luminated, but testis was not. In 35 day-old mouse, both of testis and epididymis were luminated and eaithdymal epithelium was differentiated into principal cells and clear cells. Spermatozoa were not transfered into epididymis yet. However, in 80 day-old mouse, both of festis and epididymis were fully differentiated and spermatozoa were transfered into epididymis. In electrophoretic paftem of proteins in epididymal fluid, a total of 28 kinds of proteins were identified, which were different from those of their sera. 12 kinds out of these proteins were epididymal specific protein(ESP) detected in epididymal fluid only, and the other 16 kinds(TEP) were also detected in testicular fluid. The proteins in epididymal fluid changed during sexual maturation and 3 kinds of the proteins changed quantitatively according to epididymal regions in adult. It may be concluded from the above results that the component and composition of the proteins in epididymal fluid changed by the influx of testicular fluid including spermatozoa into epididymis and regulation of the protein synthesis, secretion and/or absorption by the epididymal epithelium. Therefore it is strongly suggested that ESP and TEP in epididymal fluid play somehow significant roles on the maturation of epididymal spermatozoa.

• Applied Microscopy
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• v.22 no.2
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• pp.127-140
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• 1992

This research was undertaken to determine the effects of the cyclophosphamide (CP) on the epididymal corpus of the male rat in terms of ultrastructural alteration and protein analysis by SDS-PAGE at different groups; control group, 1 week group, 3 weeks group and 5 weeks group were treated with saline (control group) or CP at doses of 20mg/kg/week, 1 time a week, respectively. In the cytoplasm of the principal cells on the epididymal corpus, the mitochondria were significantly swollen or disrupted. The lumens of rough endoplasmic reticulum (rER) were also dilated and the number of secretory vesicles and lysosomes were increased respectively. CP caused changes in protein concentrations in the corpus of epididymis after CP treatment. Total proteins of 31 to 36 species were expressed in the corpus fluid. Then the more CP was increased, the more concentration of proteins caused to decrease, synthesize or increase in epididymal corpus. In contrast to the control group, in particular 88KD and the other 8 proteins in the corpus fluid, were decreased or disappeared respectively, whereas acid phosphatase and the other 9 proteins in the corpus fluid, were increased or synthesized respectively. The other proteins are not showed distinctive difference. It is suggested that treatment with CP alters the specific cell organelles and proteins in segment of the epididymal corpus.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.27-34
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• 1997

This study aimed to evaluate the effect of seminal vesicle fluid (SVF) on the acrosome reaction (AR) occurred spontaneously or induced by $Ca^{2+}$ ionophore A23187, follicular fluid, and progesterone in mouse epididymal sperm. SVF was divided into high (MW>10 kD) and low (MW<10 kD) fractions by ultrafiltration. The low MW fraction of SVF decreased the rate of spontaneous AR, however the high MW fraction did not. It suggested that the low MW fraction of SVF might have contained decapacitation factor(s) responsible for prolonging of time need for capacitation. When sperm preincubated for 60 min in the presence of SVF, the rate of AR induced by A23187 was decreased, but prolongation of preincubation time for 120 min significantly potentiated the AR by A23187. It suggested that addition of SVF into sperm preincubation medium imposed the epididymal sperm a condition similar to ejaculation. AR induced by human follicular fluid or progesterone was also inhibited by SVF. It suggested that substance in SVF might have affected AR of mouse sperm by inhibiting the interaction between AR inducing ligands and sperm surface receptors involved in acrosomal exocytosis.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.301-305
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• 2006

The aim of this study was to investigate whether addition of porcine epididymal fluid (pEF) into culture medium during in vitro maturation influences the nuclear maturation of porcine germinal vesicle (GV) oocytes. Porcine cumulus-oocyte complexes (COCs) from follicles were cultured in tissue culture medium 199 (TCM 199) containing pEF. After 48hr of culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. The proportion of oocytes reaching at metaphase II (M II) stage was significantly (p<0.05) increased in oocytes cultured in the media supplemented with 10% pEF during in vitro maturation than in those without pEF regardless of cumulus presence or absence (54.6% vs 22.5%, 51.7% vs 24.2%). The supplementation of pEF during maturation of oocyte enhanced oocytes maturation in a dose-dependent manner in vitro. Also significant differences (p<0.05) in the percentage of MII oocytes were observed according to exposure period in pEF. Present study suggests that pEF contains a enhancing component(s) for nuclear maturation of porcine immature oocytes in vitro.

• Reproductive and Developmental Biology
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• v.33 no.3
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• pp.125-131
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• 2009

The aim of this study is to investigate the effect of porcine epididymal fluid (pEF) on in vitro-maturation and subsequent fertilization of porcine follicular oocytes. Porcine cumulus-oocytes complexes retrieved from antral follicles were cultured in tissue culture medium (TCM)-l99 supplemented with pEF of different concentrations. At 48 h after culture, development of oocytes to germinal vesicle (GV) breakdown, metaphase I, anaphase-telophase I, and metaphase II were examined Significant (p<0.05) increase in the proportion of oocytes developed to MII stage was observed in oocytes cultured in pEF-containing TCM-l99 than in oocytes cultured in pEF-free TCM-199 (46.2% vs 16.7%), which was a dose-dependent manner. Subsequently, the proportion of monospermic fertilization were significantly (p<0.05) increased in oocytes cultured in the TCM supplemented with pEF than those cultured in pEF-free TCM-199 (51.0% vs 24.1%). In the second series of experiment, the percentage of MII oocytes was significantly (p<0.05) increased after exposure of oocytes to pEF during the first 22 h period of culture than after exposure of oocytes to pEF during the next 24 h of culture, while no significant difference in the percentage of monospermy was observed. The results of this study demonstrate that pEF contains at least enhancing component(s) for nuclear maturation.

• Childhood Kidney Diseases
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• v.1 no.1
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• pp.86-90
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• 1997

The author experienced a case of $Henoch-Sch\"{o}nlein$ purpura with epididymitis in a 6-year-old boy who was admitted to our hospital due to left scrotal pain and the relapse of purpura on lower extremities for 3 days. Scrotal ultrasonography on admission revealed homogenously enlarged left epididymal head and small amount of fluid collection in left tunical space. The size of left epididymal head decreased gradually with no more evidence of fluid collection on day 7 and recovered completely on day 21. The author reports a case of $Henoch-Sch\"{o}nlein$ purpura with epididymitis with brief review of related literatures.

• Reproductive and Developmental Biology
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• v.32 no.3
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• pp.147-152
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• 2008

The aim of this study was to investigate what components of porcine epididymal fluid (pEF) influences the nuclear maturation of porcine germinal vesicle oocytes. Porcine cumulus-oocytes complexes from follicles were cultured in TCM 199 containing pEF. After 48 h cultures, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. Maturation rate of oocytes was significantly increased in media supplemented with 10% pEF during in vitro maturation (IVM) than in those without pEF. When lipid component of pEF was removed by treating n-heptane, no significant difference was observed in maturation of oocytes between n-heptane treatrment and intact pEF group. However, the proportion of oocytes reaching at metaphase II (M II) was significantly (p<0.05) decreased in the oocytes cultured in media containing trypsin-treated pEF compared to those in media with intact pEF. When porcine GV oocytes were matured in the medium supplemented with intact pEF or pEF heated at $56^{\circ}C$ and $97^{\circ}C$, rates of oocytes remained at GV stage were 11.7%, 29.4% and 42.0%, respectively. However, there were no difference in proportion of oocytes reaching at MII stage among intact pEF group and $56^{\circ}C$ group. Present study suggests that 1) pEF contains an enhancing component(s) for nuclear maturation in vitro of oocytes, 2) protein(s) of pEF may be capable to promote nuclear maturation in vitro, and 3) enhancing component for nuclear maturation may consist of two factors, which are responsible for germinal vesicle breakdown (GVBD) and promotion of MII stage.

• Reproductive and Developmental Biology
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• v.34 no.4
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• pp.275-281
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• 2010

The aim of this study was to investigate what protein(s) of porcine epididymal fluid (pEF) are able to enhance the nuclear maturation of porcine germinal vesicle (GV) oocytes in vitro. Proteins of pEF were fractionated by affinity, ion exchange, and gel filtration chromatography. Porcine cumulus-oocytes complexes (COC) from follicles were cultured in tissue culture medium (TCM 199) containing various fractions obtained by chromatography. Porcine COCs were also cultured in TCM 199 containing various meiosis inhibitors and pEF. After 24 or 48 h culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. When porcine COCs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than 80% of oocytes were unable to resume meiosis. However, porcine COCs supplemented with pEF were able to overcome the inhibitory effect of dbcAMP and Fo. Maturation rate of oocytes was significantly (p<0.05) increased in the media supplemented with cationic protein(s) during in vitro maturation than in those with anionic protein(s) (44.1% vs 20.0%). When oocytes were cultured in the TCM 199 with fractions obtained by gel filtration, the maturation rate of oocytes was significantly (p<0.05) higher in fraction 11 containing 18 kDa than other fractions. The present study suggests that 1) dbcAMP and Fo prevent the spontaneous maturation of oocyte after isolation from follicles, and that pEF contain a substance(s) that improves meiosis resumption in vitro of porcine COCs, 2) cationic 18 kDa protein(s) are responsible for promotion of Mil stage.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.105-110
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• 2002

The objective of this study was to determine the developmental competence of in vitro matured oocytes after intracytoplasmic sperm injection(ICSI) with epididymal spermatozoa. The ovaries were obtained from slaughtered small species dogs. Oocytes matured in vitro for 24 hrs were fertilized by ICSI with epididymal spermatozoa. After ICSI, one group of oocytes was activated with 2.0 mM dimethylaminopurine or 7% ethanol for 5 min. and second group was not activated. The follicular oocytes were cultured in synthetic oviductal fluid(SOF) and TCM-199 medium containing hormones and 10% FCS for 24~48 hrs in a incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. 1. Results of IVM showed that the percentage of oocytes reaching MII after 24 h and 48 hrs of incubation were significantly higher(p<0.05) after culture with 48 hrs(9/30, 30.0%) than that after culture with 24hrs(a/30, 26.7%). 2. Results of IVM showed that the percentage of oocytes reaching MII after 48 hrs of incubation were significantly higher(p<0.05) after culture with SOF media(10/30, 30.3%) than TCM-199 media (7/30, 23.3%). 3. The rate of cleavaged embryos to blastocyst obtained by ICSI treated activation oocytes was significantly higher(p<0.05) than that of nonactivation oocytes(5/16, 25.0% vs 1/13, 5.0%). 4. The rates of development of cleavaged embryos to blastocyst obtained by ICSI treated sperm of fresh, epididymal and frozen-thawed epididymal were 8/18(44.43%), 5/16(31.3%), 2/14(14.3%), respectively. and these values of frozen-thawed epididymal sperm injection were lower than fresh sperm injection.

• The Korean Journal of Zoology
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• v.33 no.1
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• pp.78-93
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• 1990

The ultrastructure of epididymal epithelium of 10, 20, 35 and 80 day-old mouse was observed to study the differentiation and function of the epithelial cells in connection with the absorption and secretion during sexual matruation. The differentiation of epididymis was divided into three phases of, 1) undiflerentiated phase until the day 20 after birth, 2) growing and differentiating phase between the day 20 and 35, and 3) maturating phase up to the adult. Each phase was closely related with the lumination of seminiferous tubule and the influx of spermatozoa within testicular fluid from testis. In adult, the ultrastructural features appeared an absorptive function in the principal cells of proximal caput epididymis, and a strong activity of protein synthesis and secretion in distal caput, corpus and cauda epididymis. Clear cells were predominantly located in corpus and cauda epidiymis, and plenty of absorption vesicles including membranous particles assumed to be the cellular residues from spermatozoa were observed at apical region. Therefore, the distribution of various cell types of epithelium and the ultrastructure even in the same type of epithelial cell, were different according to the epididymal regions.