• 한국패류학회지
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• 제27권2호
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• pp.149-157
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• 2011

Some characteristics of the formations of acrosomal vesicles during the late stage of spermatids during spermiogenesis and taxonomical charateristics of sperm morphology in male two species (Saxidomus purpurata and Meretrix petechialis) in the family Veneridae were investigated by electron microscope observations. In two species, the morphologies of the spermatozoa have the primitive type and are similar to those of other bivalves in that it contains a short midpiece with five mitochondria surrounding the centrioles. The morphologies of the sperm nuclear types of S. purpurata and M. petechialis in Veneridae have the curved cylindrical and cylinderical type, respectively. And the acrosome shapes of two species are the same cap-shape type. In particular, the axial filament is not found in the lumen of the acrosome of two species, however, subacrosomal material are observed in the subacrosomal spaces between the anterior nuclear fossa and the acrosomal vesicle of two species. The spermatozoon of S. purpurata is approximately 46-$52{\mu}m$ in length, including a curved sperm nucleus (about $3.75{\mu}m$ in length), a long acrosome (about $0.40{\mu}m$ in length),and a tail flagellum (about 45-$47{\mu}m$ long). And the spermatozoon of M. petechialis is approximately 47-$50{\mu}m$ in length including a slightly curved sperm nucleus (about $1.50{\mu}m$ in length), an acrosome (about $0.56{\mu}m$ in length) and tail flagellum (44-$48{\mu}m$ in length). In two species, the axoneme of the sperm tail flagellum of each species consists of nine pairs of microtubules at the periphery and a pair of cental doublets at the center. Therefore, the axoneme of the sperm tail flagellum shows a 9 + 2 structure. In particular, taxonomically important some charateristics of sperm morphologies of two species in the family Veneridae are acrosomal morphology of the sperm, The axial filament is not found in the acrosome as seen in a few species of the family Veneridae in the subclass Heterodonta. The acrosomal vesicle is composed of right, left basal rings and the apex part of the acrosomal vesicle. In particular, right and left basal rings show electron opaque part (region), while the apex part of the acrosomal vesicle shows electron lucent part (region). These charateristics belong to the subclass Heterodonta, unlikely a characteristic of the subclass Pteriomorphia showing all part of the acrosome being composed of electron opaque part (region). Therefore, it is easy to distinguish the families or the subclasses by the acrosomal structures. The number of mitochondria in the midpiece of the sperm of S. purpurata and M. petechialis in Veneridae are five. However, the number of mitochondria in the midpiece of the sperm in most species of Veneridae in the subclass Heterodonta are four. Therefore, the number of mitochondria of the sperm midpiece of two species are exceptionally 5, and it is only exceptional case in the species in Veneridae in the subclass Heterodonta. Except these cases, the number of mitochondria in the sperm midpiece in all families in the subclass Heterodontaare are 4, and now widely used in taxonomic analyses.

• 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.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.56-56
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• 2001

The exogenous gene transfer by intracytoplasmic sperm injection (ICSI) procedure has been recently used to produce transgenic mice and pigs. Sperm-mediated DNA transfer has the potential to markedly simplify the generation of transgenic animals. This method may serve as an alternative to the pronucleus injection of DNA for the production of transgenic pigs. Therefore, in this study, we investigated the expression of transgene after co-injection of spermatozoon or sperm head with green fluorescent protein (GFP) gene into in vitro matured porcine oocytes. Spermatozoon and sperm head, that was obtained by sonication, were treated with 0.03% Triton X-100 to remove the membrane. They were preincubated with linearized pEGFP-N1 for 1 min, and then embryos cultured NCSU23 medium for 2.5 days after co-injected of sperm and DNA. We monitored expression of GFP in embryos under epifluorescent microscope. The remove of sperm membrane did not alter the developmental competence of embryos after ICSI. At 7 days following injection, the rates of blastocysts following injection of intact sperm (15.0%), and of sperm with disrupted membrane (14.2%) were higher than that following IVF (10.0%). Porcine oocytes injected with sperm which co-cultured with DNA concentration of 1, 0.1, and 0.01 ng were 60, 65.7 and 75% and 18.5, 37.4 and 22.2% for rates of cleavage and GFP expression, respectively. In vitro matured porcine oocytes injected with sperm and isolated sperm head resulted in 69 and 59.7% of cleavage rates, respectively The rates of embryo GFP expressed did not significantly different between sperm (20.4%) and sperm head (20.0%) injection. The transgenic embryos with the clusters of positive blastomeres were observed under fluorescent microscope. Most of embryos expressed GFP gene showed mosaicism. They showed GFP expression at 1/4, 2/4 and 3/4 of blastomeres at the 4-cell stage. Among these 4-cell embryos, the expression rate of 1/4 blastomere group (54.6%) was higher than the other groups (15.3-30.7%). These results indicate that membrane disrupted sperm could attach with exogenous DNA, and that this procedure may be useful to introduce foreign gene into porcine oocytes. Therefore, our data suggest that the ICSI car be a useful tool to efficiently produce transgenic pig as well as other mammals.

• 한국발생생물학회지:발생과생식
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• 제20권3호
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• pp.247-254
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• 2016

Cytological changes of the epithelial cells according to the developmenatal phases of the seminal vesicle related to the spermatogenic stages in the testicular lobules during spermagenesis in male Neptunea (Barbitonia) cumingii (Gastropoda: Buccinidae) were investigated monthly by electron microscopical and histological observations. N. (B) cumingii is dioecious, and an internal fertilization species. The male genital organ is located near the tentacles. The spermatozoon is approximatley $50{\mu}m$ in length. The axoneme of the tail flagellum consists of nine pairs of microtubles at the periphery and one pair at the center. The process of germ cell development during spermatogenesis can be divided into five succesive stages: (1) spermatogonia, (2) primary spermatocytes, (3) secondary spermatocytes, (4) spermatids, and (5) spermatozoa. A considerable amount of spermatozoa make their appearance in the testicular lobules (or acini) and some of them are tranported from the testis towards the seminal vesicles until late July. In this study, the developmental phases of the epithelial cells of the seminal vesicles of N. (B.) cumingii could be classified into four phases: (1) S-I phase (resting), (2) S-IIphase (early accumulating), (3) S-III phase (accumulating), and (4) S-IV phase (spent). However, in case of N. (B.) arthritica cumingii, the developmental phases of the seminal vesicle were devided into three phases: (1) resting, (2) accumulating and (3) spent. Granular bodies in the inner layer of the seminal vesicles are involved in resorption of digestion of residual spermatozoa.

• 한국발생생물학회지:발생과생식
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• 제20권1호
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• pp.11-22
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• 2016

The ultrastructures of germ cells and the functions of Leydig cells and Sertoli cells during spermatogenesis in male Kareius bicoloratus (Pleuronectidae) were investigated by electron microscope observation. Each of the well-developed Leydig cells during active maturation division and before spermiation contained an ovoid vesicular nucleus, a number of smooth endoplasmic reticula, well-developed tubular or vesicular mitochondrial cristae, and several lipid droplets in the cytoplasm. It is assumed that Leydig cells are typical steroidogenic cells showing cytological characteristics associated with male steroidogenesis. No cyclic structural changes in the Leydig cells were observed through the year. However, although no clear evidence of steroidogenesis or of any transfer of nutrients from the Sertoli cells to spermatogenic cells was observed, cyclic structural changes in the Sertoli cells were observed over the year. During the period of undischarged germ cell degeneration after spermiation, the Sertoli cells evidenced a lysosomal system associated with phagocytic function in the seminiferous lobules. In this study, the Sertoli cells function in phagocytosis and the resorption of products originating from degenerating spermatids and spermatozoa after spermiation. The spermatozoon lacks an acrosome, as have been shown in all teleost fish spermatozoa. The flagellum or sperm tail of this species evidences the typical 9+2 array of microtubules.

• Fisheries and Aquatic Sciences
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• 제6권2호
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• pp.51-58
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• 2003

The gonadal development and the gametogenic cycle and the fine structure of ripe germ cells of the cultured Pacific oyster, Crassostrea gigas were investigated using oysters monthly collected from the southern coast of Korea from October 2000 to September 2001. Monthly changes in the condition index were similar to that of meat weight rate and the highest value was observed in between April and May, and the lowest value in August. The external colors of the testis and the ovary were milky white and yellowish, respectively. The spawning period of the Pacific oyster was continued from May to September, with a peak in July. The gametogenic cycle could be classified into five successive stages: multiplicative stage (December to March), growing stage (March and April), mature stage (April to June), spawning stage (June to August) and resting stage (August to January). Variety of egg yolk granules, lipid granules, mitochondria, and endoplasmic reticula were observed in cytoplasm of ripe oocyte. The spermatozoon consisted of the head, middle piece and tail; including cap-shaped acrosome with domed structure, elliptical shaped nucleus, four mitochondria, two centrioles and flagellum.

• 한국수산과학회지
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• 제31권3호
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• pp.353-358
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• 1998

Experiments were performed to find out the physico-chemical properties of milt, and morphological changes of cryopreserved spermatozoa in tiger puffer, Takifugu rubripes. The average number of sperm and spermatocrit in milt stripped were $9.81{\pm}0.34{\times}10^{10}/m{\ell}$ and $97.8{\pm}0.8$, respectively. While total lipid concentration from seminal fluid was higher than that from sperm, total protein concentration from sperm was higher than that from seminal fluid, Na and K concentrations in sperm and those in seminal fluid were similar each other, However, glucose from sperm and seminal fluid were not detectable. Spermatozoon of tiger puffer was consisted of head, middle Piece and tail. Size of head showing horseshoe shape was $0.65{\pm}0.10{\mu}m$ in diameter and $1.35{\pm}0.30{\mu}m$ in length. The head fully containing chromatin did not have acrosome. Mitochondrion in middle piece was $0.2{\mu}m$ in average diameter and flagellum showed 9+2 structure. A few of cryopreserved spermatozoa showed morphologically loose or swollen plasma membranes.

• Fisheries and Aquatic Sciences
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• 제8권1호
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• pp.17-26
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• 2005

The ultrastructure of germ cells during spermatogenesis and the structural changes in the epithelial cells of the seminal vesicle with testicular development in male Neptunea (Barbitonia) arthritica cumingii were investigated monthly based on electron microscopic and histologic observations. N. arthritica cumingii (Gastropod: Buccinidae) undergoes internal fertilization and possesses a modified type of spermatozoon, which is approximately 20$\mu$m long. The axoneme of the tail flagellum consists of nine peripheral pairs of microtubules and one central pair. Many spermatozoa occur in the acini of the testis in the ripe stage and are transported to the seminal vesicles in the accumulating phase. In males, the monthly gonadosomatic index began to increase in September and reached a maximum in February. Subsequently, it decreased rapidly after April. The testis of this species can be classified into four developmental stages: the active (August to September), ripe (October to July), copulation (April to July), and recovery (July to August) stages. Structural changes in the epithelial cells of the seminal vesicles of this species could be classified into three phases: (1) S-I (resting), (2) S-II (accumulating), and (3) S-III (spent) phases. The morphology and structure of the epithelial cells of the seminal vesicle differed in each phase; the cells were cuboidal, squamous, or columnar in the resting, accumulating, or spent phases, respectively.

• 한국어류학회지
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• 제8권1호
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• pp.1-8
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• 1996

저서성 어류인 Cobitis striata의 정소는 좌우 한쌍의 가늘고 긴 촛대모양으로서 체벽에 매달려 있다. 또한 미성숙한 정소는 규칙적인 배열을 하는 많은 정소엽으로 구성되어 있으나 성숙된 정소에서 는 복잡한 망상구조(network)를 보였다. C. striata의 정자는 첨체가 없어 수중으로 배정되는 무첨체 수중형(anacrosomal aquasperm)이며, 구형의 핵을 갖는다. 중편은 길이가 약 $0.8{\mu}m$으로서 아주 짧았으며, ring모양의 미토콘드리아가 5~8개 존재하였다. 편모는 하나로서 전형적인 9+2구조를 보이며 돌기물(fin)이 없는 특징을 보였다.

• Asian-Australasian Journal of Animal Sciences
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• 제8권5호
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• pp.415-418
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• 1995

Semen characteristics were examined to find the deterioration of the percentage of live spermatozoa with intact acrosome during hot season using 5 Holstein bulls located in Shirnizu-cho Hokkaido Japan. Spermatozoal viability and acrosomal status were observed simultaneously with triple-stain technique for each spermatozoon. Spermatozoa were divided in four categories (live spermatozoa with intact acrosome, live spermatozoa without intact acrosome, dead spermatozoa with intact acrosome and dead spermatozoa without intact acrosome). Bull and collection month had significant effects on semen characteristics (p < 0.01). The percentage of live spermatozoa with intact acrosome and the percentage of live spermatozoa had the lowest least squares mean by collection month in August (72.7% and 76.7%). These two characteristics indicated the obvious deterioration during hot season. But the fluctuation of these two characteristics were not parallel and the differences between the two characteristics were largest during July to September. The present results indicate the necessity for the simultaneous determination of viability and acrosomal status of each Holstein bull's spermatozoa in order to keep fertility above an acceptable minimum level during hot season.