• 한국패류학회지
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• 제26권3호
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• pp.235-244
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• 2010

Ultrastructural characteristics of the testis and spermatogenesis of Crassostrea gigas were investigated by Transmission and Scanning Electron microscope observations. The testis is a diffuse organ consisting of branching acini containing differentiating germ cells in a variety of stages. The acinus is surrounded by an intermitent layer of myoepithelial cells andis divided into subcompartments that are partially separated by pleomorphic accessory cells which remain in close contact with germ cells until late stages of development. these accessory cells contain a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in the supplying of the nutrients for germ cell development, while any phenomena associated with phagocytosis of undischarged, residual sperms by lysosomes could be find in the cytoplasm of the accessory cells. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves. Mature spermatozoa consist of broad, cap-shaped acrosomal vesicle, subacrosomal material (containing axial rod embedded in a granular matrix), a oval nucleus showing deeply invaginated anteriorly, two triplet substructure centrioles surrounded by four spherical mitochondria, and satelite fibres appear to the distal centriole and plasma membrane. Spermatozoa of C. gigas resemble to those of other investigated ostreids. In particular, the anterior region of the acrosomal vesicle is transversely banded. 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 morphology in the resolution of taxonomic relationships within the Ostreidea. The spermatozoon is approximately $42-47{\mu}m$ in length including an oval sperm nucleus (about $0.91{\mu}m$ in length), an acrosome (about $0.42{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9 + 2 structure. These morphological charateristics of acrosomal vesicle belong to the family Ostreidae in the subclass Pteriomorphia.

• 한국발생생물학회지:발생과생식
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• 제13권2호
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• pp.115-122
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• 2009

기수산 2배체 일본재첩(Corbicula japonica)의 정자형성과정 및 정자의 미세 구조적 특징을 전자현미경 관찰에 의해 조사하였다. 세포학적 조사 결과, 기수산 2배체인 일본재첩의 정자 길이는 약 55${\mu}m$이다. 정자 두부(길이 약 12${\mu}m$)는 길게 신장되어 있으며 약간 구부러져 있다. 정핵 길이는 7.90 ${\mu}m$, 첨체 길이는 약 2.70 ${\mu}m$이다. 정자의 핵과 첨체의 형태는 각각 긴 화살 모양과 길다란 원추 모양을 나타낸다. 본 종(체외수정, 자웅이체, 난생종)의 정자 두부는 이미 몇몇 저자들에 의해서 보고된 3배체 재첩류(체내수정, 자웅동체, 난태생종)의 정자 두부에서 나타나는, 원시형으로부터 부분적으로 변형된 형태를 나타내고 있다. 그러나 부분적으로 변형된 2개의 편모가 있는 정자를 가지는 담수산 3배체인 자웅동체 조개류와 달리 한 개의 편모를 갖는 정자를 본 종은 생산한다. 2배체 일본재첩은 중심체를 둘러싸는 4개의 미토콘드리아를 가지고 있어, 짧은 중편을 가지는 다른 이매패류의 것들과 유사하다. 정자 미부 편모의 약소님은 중앙에 1쌍의 미세소관과 주변에 9쌍의 미세소관으로 구성되어 있다. 정자 미부의 악소님은 9+2구조를 가지며, 횡절단된 한 개의 편모를 갖는 정자에서 특히, 체외수정 어류들에서 나타는 날개 모야으이 악소님 lateral fin들이 관찰되었다.

• Applied Microscopy
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• 제33권4호
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• pp.267-274
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• 2003

난태생 어류인 성숙한 Xiphophorus maculatus의 정소를 적출하여 정자형성과정과 정자의 미세구조를 전자현미경으로 관찰하였다. 정자형성과정은 정소낭(testicular cyst)에서 이루어지며, 각 정소낭 내에 동일한 분화시기의 생식세포가 분포하고 있었다. 정원세포는 타원형으로 핵 내에 뚜렷한 인을 보유하고 있었고 세포질에는 미토콘드리아의 발달이 현저하였다. 제1정모세포는 원형으로 정원세포보다 크기가 작았고 인은 발달되어 있지 않았으며, 제2정모세포는 제1정모세포보다 더 작아졌고 핵의 전자밀도는 더 높았다. 정세포의 초기발달시기에는 세포의 크기가 정모세포보다 작았고 염색질의 응축은 핵막 쪽에서 뚜렷하여 전자밀도가 높았으며 편모가 형성되기 시작하였고 미토콘드리아는 핵 주변에 분포하였다. 정자완성과정 중기에는 핵의 염색질 응축이 뚜렷하였으며 핵은 세포질 한쪽에 치우쳐 있었고, 미토콘드리아는 편모 주변에 집중되었으며 핵은 구형을 이루고 있었다. 완전히 성숙한 정자의 두부형태는 장원추형이었고 두부에서 첨체는 관찰되지 않았으며 편모의 미세소관 배열은 9+2구조를 이루고 있었다. 또한 정자의 꼬리 끝에는 고리형태의 구조물 보유하고 있었다.

• 대한수의학회지
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• 제37권4호
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• pp.925-934
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• 1997

This study was directed at inducing the production of antibodies by immunizing heifers with bovine sperm antigen and on measuring the serum antibodies using indirect immunofluorescence assay(IFA) and agglutination test. The effect of antisperm antibodies on fertilizing capacity of bovine spermatozoa was evaluated. 1. Three heifers between 12- and 15- month old were immunized with bovine spermatozoa or phosphate-buffered saline. In heifers immunized with bovine spermatozoa serum IgG level was highest between 3 weeks and 5 weeks postimmunization detected by IFA. The antibody levels persisted through week 7 and slowly declined until week 20 and then antisperm antibodies were localized on spermatozoa. The fluorescent antisperm antibodies were detected at 2~20 weeks and at 6~9 weeks postinoculation on acrosome and tail, respectively. Among 21 sera from repeat breeder cows, only one cow has shown positive antisperm antibody response detected by IFA. 2. In spite of vital rate of bovine sperm after swim-up was not significantly affected by different concentration of antisperm antibodies in sera, the numbers of bovine sperm after swim-up were significantly reduced in proportion to the increased concentration of antibodies. Above 1/512 dilution of antibody neither influence on vital rate and numbers of bovine sperm nor sperm agglutination after swim-up. The study has also shown that the vital rate and number of sperm after swim-up and capacitation were also significantly reduced by the addition of antisperm antibodies. Although antisperm antibodies did not influence on the acrosome reaction rate of sperm during swim-up, did significantly reduce the sperm acrosome reaction rate after capacitation. The studies have resulted that the bovine antisperm antibodies can prevent the sperm motility by agglutination and block the capacitation and acrosome reaction of bovine sperm.

• 한국가축번식학회지
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• 제17권1호
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• pp.1-6
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• 1993

This experiment was conducted to determine the effects of gossypol injection spermatogenesis of mice. Gossypol was injected into the stroma of testes(TS) and the doses of gossypol injected were 5, 10 and 15mg per kg of body weights, respectively. The number of sperm and the weight of testes were gradually reduced(P<0.01) from 2 to 4 weeks after gossypol treatment in all groups of mice treated with gossypol, compared with the control group. The rates of malformation(loss of proacrosome, damage of midpiece and breaking of tail) of sperm were significantly(P<0.01) increased at 2 and 3 weeks after the injection of 10 or 15mg of gossypol. However, the weight of testes and the number of normal sperm were gradually increased and the malformation rate of sperm was decreased between 4 and 6 weeks after injection of 5mg of gossypol. The results of this experiment indicated that probably ireeversible suppression of spermatogenesis could be brought about easily and immediately by the single injection of gossypol into TS.

• Reproductive and Developmental Biology
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• 제29권3호
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• pp.149-154
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• 2005

The purpose of this study was the analysis of sperm ability in Specific Pathogen Free (SPE) miniature pig for production of bio-organ. The collected semen was diluted with extender and stored at $17^{\circ}C$t for up to 7 days. The semen samples were evaluated at 0, 1, 3, 5, and 7 days of storage for analysis of sperm ability. Sperm ability was evaluated by examining viability, progressive motility, sperm abnormality and intensity of the sperm membrane. Also, the semen was processed according to the convenient freezing method, and frozen-thawed sperm was evaluated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) staining. Motility of spermatozoa of SPF miniature pig was significantly (P<0.05) lower on 3 days or later compared to the Duroc, Yorkshire and Landrace in domestic boar. The percentage of abnormal spermatozoa of Landrace were significantly (P<0.05) higher than in SPF miniature pig, Duroc and Yorkshire that had a similar percentage on 5 or 7 days of sperm storage. The percentage of spermatozoa with coiled tail decreased during the storage period but there were no significant difference. On the other hand, viability of frozen-thawed spermatozoa had a significantly (P<0.05) lower in SPF miniature pig than in other domestic boars. CTC patterns had no significant difference, but SPF miniature pig had higher percentage of capacitated spermatozoa and lower percentage of acrosome-reacted it than domestic boars. Therefore, this study suggest that it is necessary to develop the suitable extender and freezing methods methods for the high viable rate and fertilizing ability in vitro.

• 한국수정란이식학회지
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• 제31권4호
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• pp.355-365
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• 2016

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

• 한국발생생물학회지:발생과생식
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• 제18권3호
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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.

• Applied Microscopy
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• 제39권2호
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• pp.89-99
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• 2009

Cricetinae 아과 Clethrionomys 아속에 속하는 한국산 대륙밭쥐(Clethrionomys rufocanus)의 정자의 형태적 특징을 알아보기 위하여 주사 및 투과형전자현미경으로 관찰하였다. C. rufocanus정자머리의 모양은 날카롭게 굽은 하나의 dorsal hook를 가지며, 전체적인 머리모양은 마치 도끼모양의 형태를 취하고 있었다. C. rufocanus정자의 전체길이는 95.8 ${\mu}m$이었으며, 정자머리의 길이는 7.8 ${\mu}m$, 꼬리는 88.0 ${\mu}m$이었다. 꼬리는 경부 (1.0 ${\mu}m$), 중편부(22.0 ${\mu}m$), 주편부와 미부 (65.0 ${\mu}m$)로 각각 구성되어져 있었다. 분절주 (segmented columns)는 약 10~12개의 마디로 이루어져 있었으며, 미토콘드리아의 수는 약 170~178개 정도이었다. 첨체 후 덮개는 핵의 1/2을 점유하고 있었으며, 적도절은 핵 표면의 첨체 후 덮개(post-nuclear cap)와 첨체덮개(acrosomal cap) 사이에 위치하고 있었다. 1, 5, 6번의 외측섬유들은 다른 것보다 더 크며, 섬유초 (fibrous sheath)와 종 지주(longitudinal column)은 주편부에서 관찰되어졌으며, 미부에서는 섬유초가 존재하지 않았다. 이상의 결과로 미루어 보아, 정자두부와 꼬리의 형태적인 구조는 정자의 진화 패턴과 종을 분류하는데 유용한 정보를 제공하리라 여겨진다.

• 한국발생생물학회지:발생과생식
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• 제23권1호
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• pp.11-19
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• 2019

The study was conducted to investigate the effects of alpha-linolenic acid (ALA) combined with bovine serum albumin (BSA) or methyl-beta-cyclodextrin (MBCD) on plasma and acrosomal membrane damages, mitochondrial activity, morphological abnormality, motility, and oxidative stress in frozen-thawed boar sperm. In previous our study, 3 ng/mL ALA had been shown protective effect during freezing process of boar sperm. Therefore, we used 3 ng/mL ALA in present study and ALA was combined with same molar ratio of BSA or MBCD (ALA+BSA and ALA+MBCD, respectively). To confirm the effect of two carrier proteins, same volume of BSA and MBCD without ALA were added during cryopreservation. Membrane damage, mitochondrial activity, reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were measured using flow cytometry, and movement of sperm tail as motility parameter and morphological abnormality were observed under light microscope. In results, all of sperm parameters were enhanced by ALA combined with BSA or MBCD compared to control groups (p<0.05). Mitochondrial activity, morphological abnormality, ROS and LPO levels in ALA+BSA or MBCD groups were no significant difference compared with ALA, BSA and MBCD treatment groups. On the other hand, plasma and acrosomal membrane intact, and sperm motility in ALA+MBCD group were higher than single treatment groups (p<0.05), whereas ALA+BSA did not differ. Our findings indicate that carrier proteins such as BSA and MBCD could improve the effect of ALA during cryopreservation of boar sperm, and treatment of ALA with carrier proteins enhance membrane integrity, mitochondrial activity through reduction of ROS-induced LPO.