• Journal of Marine Life Science
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• v.8 no.2
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• pp.115-120
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• 2023

This study aims to find out a suitable extender and cryoprotective agent (CPA) for cryopreservation and its optimum concentration in order to conduct planned artificial seed production of Korean bullhead Pseudobagrus fulvidraco and to preserve superior sperm. Experiments were designed to investigate the effects of the different combinations of three extenders (I: 300 mM glycose, II: Kurokura extender, III: Li extender), four cryoprotectants (dimethyl sulfoxide, ethylene glycol, methanol and glycerol) and four concentrations (5, 10, 15, 20%) on the cryopreservation of Korean bullhead sperm. Postthawed sperm survival rate and sperm activity index (SAI) were detected to evaluate the effects of sperm cryopreservation. The optimal combination of extender and CPA for cryopreservation of Korean bullhead sperm was extender III + 10 and 15% methanol, resulting in a survival rate and SAI of 66.9 ± 8.7, 67.3 ± 13.1% and 2.6 ± 0.4, 2.6 ± 0.5 respectively, which was higher than had been achieved with other extenders and CPAs.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.1
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• pp.83-88
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• 2010

Objective: Globozoospermia is observed in <0.1% of male infertile patients. The aim of this study is to report the successful clinical outcome in man with globozoospermia. Design: A case report. Method: A 36-year-old man and 32-year-old partner visited our fertility center with 6 years infertility duration. The semen analysis revealed globozoospermia combined with oligozoospermia which was stained with Diff-Quick method. ICSI (Intracytoplasmic sperm injection) was performed on 12 matured oocytes. Result: 5 injected oocytes were fertilized normally. 5 embryos were transferred on day 3 after oocyte retrieval. The patient became pregnant and delivered a healthy boy at 39 weeks of gestation. Conclusion: In case of globozoospermia, it has a low fertilization rate even though ICSI method is used. The favorable technique is still needed to increase the fertilization rate.

• Journal of Biomedical Engineering Research
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• v.17 no.1
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• pp.25-32
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• 1996

A new analytic method has been developed for the analysis of sperm morphology using Hough transform. This method is based on the characteristic that sperm heads have elliptic shape in addition to the density difference with the background Sperm heads are represented in elliptic form with five parameter, and the optimal parameters are estimated by iterative Hough transform. To reduce processing time practically, we restricted the transformed space in minimum volume and moved the searching volume to the maximum gradient for the estimated error. Morphological parameters were calculated from estimated sperm head boundaries without further processing.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2002.10a
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• pp.241-242
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• 2002

수서동물 생식소의 해부학적 구조 및 생식세포의 미세구조는 종 특이성, 생식생태 및 계통에 따라 다르다. 일반적으로 조개류의 정자는 핵과 첨체의 모양, 첨체기둥(acrosomal rod)의 유무, 미토콘드리아 수, 편모의 수 및 형태 등에서 다양한 특징을 보인다 (Morse and Zardus, 1997). Anadara trapezia 등이 속하는 돌조개과 (Arcidae) 조개들의 정자 두부는 난형이며, 첨체기둥이 없고, 중편에 4개의 미토콘드리아를 가진다. (중략)

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.167-172
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• 2004

This study was performed to examine the correlations among dog sperm viabilities evaluated by flow cytometry, by microscopic evaluation (ME), by carbo-xifluorescein diacetate and propidium iodide (CFDA/PI) and by hypoosmotic swelling (HOS) test. Semen were collected from 5 dogs ranging in age from 2 to 4 years. Each ejaculate was divided into 3 aliquots and different proportions of freeze-killed cells were added to each aliquot (1:0, 1:1 and 1:3). In the other experiment, semen was extended with Sweden extender containing 5% glycerol and equex STM paste, and frozen using liquid nitrogen vapor. Fresh and frozen-thawed dog sperm viability were assessed by flow cytometry using PI staining method. The accuracy of flow cytometry was evaluated by comparing with other classic assessments, microscopic evaluation, epifluorescence microscopic analysis using CFDA/PI, and HOS test. High correlations of sperm viabilities were found among flow cytometry, epifluorescence evaluation, HOS test (p<0.01) in fresh semen. Especially, sperm viability assessed by HOS test was highly correlated with viability by flow cytometry in all the ratios of live and dead spermatozoa, 1:0, 1:1 and 1:3 (p<0.01). The viability evaluated by ME were significantly correlated with that by flow cytometry in ratios of 1:0 and 1:3 (p<0.05) however, there was no significance in ratio of 1:1. The viability evaluated by C/p were highly correlated with that by flow cytometry in ratio of 1:0 and 1:1 (p<0.01) and significantly correlated in ratio of 1:3 (p<0.05). In frozen-thawed spermatozoa, the viability determined by HOS test was considerably correlated with that by flow cytometry (p<0.01). There was significant correlation between the viabilities by ME and by flow cytometry (p<0.05). But the viability evaluated by CFDA/PI was not correlated with viability by flow cytometry. The result from this study validate the use of flow cytometry as a precise method for assessing the viability of fresh and frozen-thawed dog spermatozoa.

• Applied Microscopy
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• v.41 no.2
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• pp.99-107
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• 2011

Light microscope, and scanning and transmission electron microscopy were used to investigate the fine structure of sperm of the Korea squirrel, Tamias sibiricus. The sperm head of T. sibiricus was paddle in shape. The total length of T. sibiricus sperm was 67.8 ${\mu}m$. The length of sperm head was 7.8 ${\mu}m$, and the tail (60.0 ${\mu}m$) was consisted of four major segments: the neck (1.0 ${\mu}m$), middle (8.0 ${\mu}m$), principal (48.5 ${\mu}m$) and end piece (2.5 ${\mu}m$), respectively. Especially, the length of the middle piece is short, and end piece was very shorter than those of other rodents. The post-nuclear cap was occupied about a fifth of nucleus. The equatorial segment is located between the post-nuclear cap segment and acrosomal cap on the nuclear surface. Nine segmented columns were surrounded by the mitochondria, and numbers of gyres of mitochondria were 26. One segmented column was consisted ten to twelve knobs, and each of segmented column in the neck region connected with the nine outer dense fiber in the middle piece. Numerous satellite-like fibers were scattered around the segmented columns. Nos. 1, 5 and 6 of the outer dense fibers in the middle piece were larger than the others. A fibrous sheath and longitudinal column of the principal piece were in evidence, but the fibrous sheath and longitudinal column was not seen at the end piece. In conclusion, the structural features of sperm head and tail may be useful information to patterns of sperm evolution and classification of species.

• Journal of The Korean Society of Grassland and Forage Science
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• v.38 no.4
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• pp.320-324
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• 2018

In this study, we examined effect of sperm penetration of oocytes after in vitro fertilization (IVF) with cauda epididymal spermatozoa in Hanwoo bull after feeding of timothy hay. One testicle with epididymides was castrated from one Hanwoo bull (14 months of age) and spermatozoa recovered from cauda epididymis and cryopreserved. As control, frozen Hanwoo semen was used. Matured cumulus oocyte complexes were co-incubated with frozen-thawed cauda epididymal spermatozoa for 12 or 18 hours. After IVF, presumptive zygotes were cultured in modified synthetic oviductal fluid. In experiment 1, we examined sperm penetration rate at 12 hours of IVF with epididymal sperm. Total penetration rate among cauda epididymis and control was similar(mean${\pm}$standard error, cauda epididymis and control vs. $49.7{\pm}11.3$ and $54.4{\pm}12.8%$). In experiment 2, cleavage and blastocyst developmental rate were evaluated at day 2 and day 8 after IVF for 18 hours. Cleavage rate among cauda epididymis and control was similar(cauda epididymis and control vs. $81.2{\pm}3.4$ and $82.7{\pm}2.5%$). However, blastocyst developmental rate of cauda epididymis group was significantly higher than that of control group(cauda epididymis and control vs. $24.4{\pm}1.6$ and $12.2{\pm}2.8%$, p<0.05). In conclusion, cauda epididymal spermatozoa in Hanwoo bull has high embryo developmental competence and can be used as an alternative to ejaculated frozen sperm in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.2
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• pp.179-185
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• 1989

불염여성에서 복강내액의 독성 유무를 알기 위해서 콤퓨터 정액 분석기를 이용하여 복강내액의 정자 활동성을 분석하였다. 연구대상은 자궁내막증 환자 14예와 자궁내막증이 없는 불염환자로부터의 복강내액 9예로 하였다. 연구방법은 건강인의 정자를 셰척 분리하여 대상환자의 복강내액과 Hams F10 배양액을 동량 흔합하여 (50%) 배양후 정자 활동성을 측정하였다. 정자 활동성의 감소는 배양전 0시간의 수치와 비교해서 1시간， 4시간 그리고 24시간후의 결과를 비교 분석하였다. 자궁내막증 제 1기 및 제 2기 환자와 자궁내막증이 없는 환자의 복수에서의 정자 활동성의 감소는 통계학적으로 의의있는 차이는 없었다. 그러나 자궁내막증이 심한 제 3기의 환자군에서는 의의있는 차이가 있었다(p<.05). 실험군간의 정자 활동성의 평균치 차이는 없었으나，각 군에서의 환자별 개인 성적은 정자 활동성이 현저히 감소한 독성있는 복강내액과 비독성의 복강내액을 쉽게 구별할 수 있었다. 결론적으로 정자 배양을 이용한 복강내액 독성 검사는 비교적 간단하고 경제적이므로 염상적으로 최근에 대두된 복강내 수정 치료법의 예비검사로서 유용할 것으로 사료된다.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.08a
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• pp.36-37
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• 2001

큰가리비는 자웅이체이다. 난환형성과정은 난모세포의 발달정도에 다라 다르게 나타나고 있다. 난자형성과정은 난원세포기, 전난황형성난모세포기, 초기난황형성난모세포기, 후기난황형성난모세포기, 성숙난모세포기의 연속적인 5단계의 과정으로 나눌 수 있었다. 전난황형성기 난모세포질 내에서는 핵주변 구역에 골지장치와 수많은 공포들 및 미토콘드리아들이 출현하고 있는데 이들은 차후, 지방적 형성에 관여한다. 난황형성전기난모세포(previtellogenic oocyte)에서는 지방적 및 지질과립들이 핵막 근처에서 출현하여 피질층쪽으로 분산되는 반면, 같은 발달 단계의 난모세포질의 피질구역에서는 피질과립들(단백질성 난황과립)이 처음으로 생성되어 난황막 근처의 피질층에서 핵주변 구역쪽으로 분산.분포된다. 난황형성후기 난모세포에서는 세포질 내의 골지장치, 공포, 미토콘드리아, 그리고 조면소포체들이 자율합성에 의해 난황과립 형성에 관여하고 있다. 반면 외인성 물질들인 지질형태의 과립들, 다량의 글리코겐 입자들이 생식상피 내에서 출현하고 있는데. 이들 물질이 생식상피에서 난황막 구조물인 미세융모를 통해 난황형성 후기 난모세포의 날질 내로 통과해 들어가는 현상이 관찰되었다. 이와같은 현상은 난황형성이 일어날 때에 heterosynthesis가 일어나고 있음을 시사한다. 완숙난모세포의 난경은 약 50~60$\mu\textrm{m}$이다. 정자형성과정은 정원세포기, 제1차정모세포기, 제2차정모세포기, 정세포기, 정자기의 연속적인 5단계로 나눌수 있었다. 정셍포기에서 정자로 변태되는 과정 중에 침체의 분화과정이 있는데 이에는 1. Golgi phase, 2. Cap phase 3. acrosome phase, 4. maturation phase의 단계를 거쳐 첨체가 완성된다. 정자는 원시적 형태를 이루고 있으며 4개의 미토콘드리아가 부핵을 형성하고 있다. 완숙정자 두부의 길이는 대략 $3 \mu$m 이며, 미부의 길이는 약 $30 \mu$m정도이다. 정자 미부편모의 axoneme은 중앙의 2개의 미세소관(microtubule)과 주변에 위치한 9개의 2중 미세소구관(microtublue)으로 이루어져 있다.

• Korean Journal of Ichthyology
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• v.21 no.2
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• pp.81-86
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• 2009

The spermiogenesis and mature spermatozoa of Pseudobagrus fulvidraco were described by means of scanning and transmission electron microscopy. Spermiogenesis is characterized by lateral development of the flagellum, nuclear rotation, deep nuclear fossa formation and compaction into thick granules. The spermatozoa exhibit a round head containing a nucleus that lacks an acrosome, and having a midpiece and a flagellum. The midpiece is small and has a short cytoplasm including several mitochondria separated from the tail by the cytoplasmic canal. The flagellum contains the 9+2 classical axoneme structure and has two axonemal fins. The presence of axonemal fins in the flagellum is a common character in Bagridae. The interrelationships among the Bagridae as well as other teleosts are herein discussed.