• Title/Summary/Keyword: Sperms

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Microtubule Assemblies and Methylation Patterns of Porcine IVF and Parthenogenetic Embryos (돼지 체외수정란 및 단위발생란의 미세관 형성 및 메틸화 양상)

  • Park, Joo-Hee;Kim, Ho-Jeong;Lee, Beom-Ki;Kwon, Dae-Jin;Hwang, In-Sun;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.7-11
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    • 2009
  • This study was performed to confirm the microtubule assemblies and methylation patterns of porcine IVF and parthenogenetic embryos. Cumulus-oocyte complexes were collected and matured in vitro for 42 hr. Oocytes were fertilized by prepared fresh sperm or activated parthenogenetically by exposure to electric stimulation and 6-dimethylaminopurine. Porcine IVF and parthenogenetic embryos were cultured in vitro for 6 days. Embryos were stained by immunofluorescence staining method to observe the dynamic of nucleus and microtubules in the first mitotic phase and the methylation patterns in different developmental stages. After then, samples were confirmed and analyzed through a laser-scanning confocal microscope. IVF embryos had a centrosome originated from sperms, which was shown a $\gamma$-tubulin spot. However, $\gamma$-tubulin spot was not observed in parthenogenetic embryos. A lower methylation level was observed in IVF embryos compared to parthenogenetic ones at the morula and blastocyst stages. In conclusion, it is considered that microtubule assemblies and genetic regulation mechanism differ between parthenogenetic and IVF embryos.

Effect of an Anabolic Steroid, Nandrolone Decanoate, on Aquaporin 1 and 9 Gene Expression in the Rat Epididymis

  • Seo, Hee-Jung;Kang, Hyo-Jin;Choi, In-Ho;Cheon, Yong-Pil;Lee, Ki-Ho
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.55-61
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    • 2009
  • The epididymis in the male reproductive tract is the site where spermatozoa produced from the testis become mature. The epididymis is divided into 4 different segments, initial segment and caput, corpus, and caudal epididymis, depending upon functional and morphological features. Aquaporins (Aqps) are water channel molecules, which are present in the epididymis and play a major role in removal of epididymal water, resulting in creation of microenvironment for sperm maturation and concentration of sperms. Nandrolone decanoate (ND) is a synthetic anabolic-androgenic steroid, which is used to treat clinical diseases and improve physical ability and appearance. Even though it is well determined that the ND causes the male infertility by affecting the testis, little is known the effect of the ND on the epididymis. The present study was focused to examine the effect of ND at different treatment doses and periods on expression of Aqp1 and Aqp9 genes in the epididymis of pubertal rats. Results showed that mRNA expression of Aqp1 and Aqp9 genes among the parts of the epididymis was differentially regulated by ND treatment doses. In addition, treatment periods of ND caused differential expression of Aqp1 and Aqp9 mRNAs among segments of the epididymis. Therefore, it is believed that male infertility induced by ND could be resulted not only from malfunction of the testis but also from aberrant gene expression of Aqp1 and Aqp9 in the epididymis.

Analysis of Membrane Integrity, DNA Fragmentation and Mitochondrial Function in Pig Spermatozoa Sorted by Flowcytometer

  • Kim, In-Cheul;Han, Deug-Woo;Lee, Sung-Won;Ryu, Jae-Weon;Choi, Eun-Ji;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • v.32 no.2
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    • pp.123-126
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    • 2008
  • The objective of this study was to determine the potential hazardous effects of sorting process by flowcytometry on the quality of boar spermatozoa by flowcytometer. Freshly collected boar semen was diluted and divided into two groups; control none sorted and sorted. Sperms in sorted group were processed with flowcytometer for cell sorting with $100\;{\mu}M$ nozzle under the 20 psi pressure. Measurements on each parameter were made at two time points, 0hr (right after sorting) and 24 hr post sorting. Although there was a tendency of lower viability in sorted group than none sorted control group, the percentage of live cells in control ($75.83{\pm}6.92\;&\;59.53{\pm}10.34$) was not significantly different from sorted ($59.70{\pm}7.37\;&\;43.97{\pm}3.76$) at both 0 and 24 hr post sorting. However, sorted sperm showed significantly lower mitochondrial function compared to the control at both 0 h ($79.37{\pm}3.22\;vs.\;63.50{\pm}10.05$) and 24 hr ($67.27{\pm}3.22$ vs. $46.97{\pm}5.37$) time points (p<0.007). Sperm DNA fragmentation rate was significantly lower in control ($22.0{\pm}7.04$) than that of sorted ($32.27{\pm}7.49$) at 24 hr time point (p<0.0002). Taken together, these data suggested thatsorting process by flowcytometer may have influenced sperm motility rather than viability. Also high speed sperm sorting by flowcytometer has significant effects on DNA fragmentation on elapsed time after sorting.

Effects of α-Linolenic Acid and Bovine Serum Albumin on Frozen-thawed Boar Sperm Quality during Cryopreservation

  • Lee, Won-Hee;Hwangbo, Yong;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.40 no.4
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    • pp.33-37
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    • 2016
  • This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

First observation on the early embryonic and larval development of spiny oyster Saccostrea kegaki Torigoe & Inaba, 1981 (Bivalvial: Ostreoida) using scanning electron microscope on the north coast of Jeju, Korea (주사전자현미경 (Scanning Electron Microscope)을 이용한 제주 북부 연안에 서식하는 가시굴 (Saccostrea kegaki Torigoe & Inaba, 1981)의 초기 유생발달관찰)

  • Lee, Hee-Jung;Kang, Hyun-Sil;Jeung, Hee-Do;Hong, Hyun-Ki;Choi, Kwang-Sik.
    • The Korean Journal of Malacology
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    • v.29 no.2
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    • pp.97-103
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    • 2013
  • In the present study, we monitored the early development of Saccostrea kegakia subtropical oyster species distributing on rocky intertidal off the northern Jeju Island using scanning electron microscope (SEM). The female oyster collected in early August, 2012 were fully mature exhibiting relatively small eggs ($46.5{\pm}1.4{\mu}m$ in diameter) in the gonad, while testis of the mature male oysters were filled with fully developed sperms of 36.9 ${\mu}m$ in length. The fertilized eggs developed into 2-cell stage with polar body after 1 hr 20 min of fertilization, then followed by Morula stage (3 hr 20 min), Blastula stage (4 hr 50 min), Gastrula stage (7 hr), and trochophore larvae stage (9 hr 30 min). The observed early development of S. kegaki in this study was similar the early development of other oysters, although size of the fertilized eggs were somewhat smaller.

A Study on the Isolation of Ureaplasma urealyticum in Nongonococcal Urethritis (비임균성 요도염에 있어서 Ureaplasma Urealyticum의 분리에 관한 연구)

  • Lee, Young-Tae;Lee, Moo-Sang
    • Clinical and Experimental Reproductive Medicine
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    • v.10 no.1
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    • pp.25-37
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    • 1983
  • A group of 180 men who visited Urology Department of Severance hospital, including 115 patients with nongonococcal urethritis (N.G.U.), 27 patients with prostatitis, 13 patients with gonococcal urethritis (G.U.) and 25 healthy medical student controls were investigated for the isolation of Ureaplasma urealyticum (T-strain mycoplasma) from the specimen of ureaplasma discharge, urine and semen. Taylor-Robinson media of T-broth and T-agar was used for the isolation of Ureaplasma urealyticum. To the best of our knowledge, the study on the culture of Ureaplasma urealy ticum was reported for the first time in Korea. The followis g results were obtained: 1. The isolation rate of Ureaplasma urealyticum in nongonococcal urethritis (53.0%) revealed highest of those in the other three groups of prostatitis, gonococcal urethritis and control (40.7%,38.4% and 16.0% respectively). 2. As for the specimens, urethral discharge revelaed higher isolation rate of Ureaplasma urealyticum (54.6%) than first voided urine (50.0%). 3. The more consorts patients had, the higher positive culture rate of Ureaplasma urealyticum were revealed. The isolation rate in case of more than one causal in nongonococcal urethritis (27.8%) revealed much higher than in case of marital only (5.2%), one regular (6.1%) and one causal 03.9%). 4. 2.6% of isolation rate of Ureaplasma urealyticum revealed in patients with nongonococcal urethritis who visited the clinic in later than 4 weeks after the symptoms developed. However, the isolation rate in patients who visited within 4 weeks revealed 50.3%. The lower isolation rate of Ureaplasma in the late treatment seekers might be probably due to the suppression effect against Ureaplama urealyticum from the possible previous self antibiotic treatment. 5. Attachment of Ureaplasma urealyticum mostly to the neck and head portion of the spermatozoa seemed to playa role to affect the motility of sperms.

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Magnetic Orientations of Bull Sperm Separated into Head and Flagellum Treated by DTT or Heparin

  • Suga, D.;Shinjo, A.;Kurnianto, E.;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.2
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    • pp.167-175
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    • 2000
  • This paper describes the magnetic orientation of bull sperm separated into the head and the flagellum treated by DTT or heparin in a 5,400G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4^{\circ}C$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the head area at 1, 3 and 6 days. After measuring the area, each sample was exposed to a 5,400G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the sperms were separated into the head and the flagellum through the DTT treatment. Almost of the separated heads showed that their long axis oriented perpendicularly to the magnetic lines of force, and most of the long axis perpendicularly oriented heads showed that their flat plane oriented perpendicularly in a 5,400G magnetic field. Also, the demembranation of the head tended to increase those perpendicular orientations, while those perpendicular orientations of the head declined with the decondensation of the sperm nuclei. These findings suggest that strong magnetic anisotropy for the perpendicular orientation of the long axis and the flat plane of the head occurs in the sperm nuclei in a 5,400G magnetic field. The separated flagellum showed lower parallel orientation, and the separated and demembranated flagellum showed parallel orientation to the magnetic lines of force in this magnetic field. These findings suggest that weak magnetic anisotropy of the parallel orientation of the flagellum occurs in the inside components in a 5,400G field.

Spermiogenesis and Taxonomical Values of Sperm Ultrastructures in Male Crassostrea ariakensis (Fujita & Wakiya, 1929) (Pteroirmorphia: Ostreidae) in the Estuary of the Seomjin River, Korea

  • Son, Pal Won;Chung, Jae Seung;Kim, Jin Hee;Kim, Sung Han;Chung, Ee-Yung
    • Development and Reproduction
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    • v.18 no.3
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    • pp.179-186
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    • 2014
  • Characteristics of the developmental stages of spermatids during spermiogenesis and phylogenetic classicfication of the species using sperm ultrastructures in male Crassostrea ariakensis were investigated by transmission electron microscope observations. The morphology of the spermatozoon of this species has a primitive type and is similar to those of Ostreidae. Ultrastructures of mature sperms are composed of broad, modified cap-shaped acrosomal vesicle and an axial rod in subacrosomal materials on an oval nucleus, four spherical mitochondria in the sperm midpiece, and satellite fibres which appear near the distal centriole. The axoneme of the sperm tail shows a 9+2 structure. Accordingly, the ultrastructural characteristics of mature sperm of C. ariakensis resemble to those of other investigated ostreids in Ostreidae in the subclass Pteriomorphia. In this study, particularly, two transverse bands (stripes) appear at the anterior region of the acrosomal vesicle of this species, unlike two or three transverse bands (stripes) in C. gigas. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm ultrastructures and morphologies in the resolution of taxonomic relationships within the Ostreidae in the subclass Pteriomorphia. These spermatozoa, which contain several ultrastructures such as acrosomal vesicle, an axial rod in the sperm head part and four mitochondria and satellite fibres in the sperm midpiece, belong to the family Ostreidae in the subclass Pteriomorphia.

Effect of L-carnitine on sperm quality during liquid storage of boar semen

  • Yang, Kang;Wang, Na;Guo, Hai-Tao;Wang, Jing-Ran;Sun, Huan-Huan;Sun, Liang-Zhen;Yue, Shun-Li;Zhou, Jia-Bo
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.11
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    • pp.1763-1769
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    • 2020
  • Objective: This study was conducted to investigate the effect of L-carnitine on the pig semen characteristics during storage. Methods: Spermatozoa samples were examined for spermatozoa quality and then randomly divided into 5 groups: 0 (control), 12.5, 25, 50, and 100 mM L-carnitine. Sperm motility, plasma membrane integrity and antioxidant parameters (total reactive oxygen species, total antioxidant capacity, and malondialdehyde) were evaluated after 0, 3, 5, and 10 day cooled-storage at 17℃. Moreover, ATP content, mitochondria activity as well as sperm-binding and in vitro fertilizing ability of preserved boar sperm were also investigated. Results: Supplementation with 50 mM L-carnitine could effectively maintain boar sperm quality parameters such as sperm motility and membrane integrity. Besides, we found that L-carnitine had positive effects on boar sperm quality mainly through improving antioxidant capacities and enhancing ATP content and mitochondria activity. Interestingly, by assessing the effect of L-carnitine on sperm fertility and developmental potential, we discovered that the extender containing L-carnitine could improve sperm quality and increase the number of sperms bounding to zona pellucida, without improving in vitro fertility and development potential. Conclusion: These findings suggested that the proper addition of L-carnitine to the semen extender improved boar sperm quality during liquid storage at 17℃.

Copulation and Embryonic Development of the Viviparous Teleost Surfperch, Ditrema temmincki (태생 경골어류, 망상어(Ditrema temmincki)의 교미 및 체내자어의 발달)

  • LEE Jung Sick;AN Cheul Min;CHIN Pyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.1
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    • pp.26-34
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    • 1996
  • Copulation process, fertilization and gestation of the viviparous teleost surfperch, Ditrema temmincki were investigated by using photomicroscopy. Samples were collected from the vicinity of Suyoung Bay, Pusan, Korea from May 1992 to August 1993. During the copulation period, the copulatory organs were protruded at the base part of right and left soft ray of the anal fin in mature male. Secondary sexual characteristics index (SSCI) of male participated in copulation was above 3.5. Mature oocytes appeared only in the female containing sperms in the ovarian cavity. Transport of the spermatozoa into the ovarian cavity during copulation belonged to a spermatophore type. After copulation, spermatozoa remained in the ovarian cavity for about one month untill fertilization. Fertilization occured within the follicular cavity. Fertilized eggs were released into the ovarian cavity where they developed during gestation period. Developmental sequence of the female was as following: fertilization-ovulation-hatching-parturition. Right before parturition, the total length (TL) of the embryo was about 63.0 mm. When TL of maternal body was 20.0 cm, the mean numbers of the embryo were 18. The numbers of the embryo were positively related to the maternal body size.

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