• Journal of Marine Life Science
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• v.6 no.1
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• pp.23-30
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• 2021

The differentiation process of male germ cells and sperm morphology of the abalone Haliotis discus hannai were described in ultrastructure. The differentiation process of sperm was divided into four stages: spermatogonium, spermatocyte, spermatid and sperm. The process of differentiation from spermatogonium to spermatocyte did not show significant morphological changes. However, during the spermiogenesis there were distinct morphological changes such as chromatin condensation, morphological changes of the nucleus, and formation of acrosome, midpiece and flagellum. The sperm of the abalone consisted of head, midpiece and tail. The head of approximately 5.3 ㎛ in length was composed of a nucleus of high electron dense and bullet-shaped acrosome. The midpiece was composed of the basal body and mitochondria, and five mitochondria were arranged in single layer around the basal body. The cross section of the tail showed a "9+2" axonemal structure. These morphological and structural features are the result of showing that the sperm of H. discus hannai is a primitive type.

• Applied Microscopy
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• v.26 no.3
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• pp.267-275
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• 1996

The internal ultrastructural changes of germ cells and external morphology of spermatozoon during the spermatogenesis in the rockfish, Sebastes inermis were studied using transmission and scanning electron microscope. The testis is seminiferous tubule type in internal structure. Seminiferous tubule consist of many cyst which contain numerous germ cells in same developmental stage. Spermatogonium contained a large nucleus with single nucleolus in interphase. Primary spermatocyte identified by the presence of synaptonemal complex in nucleus and the contained a number of mitochondria, endoplasmic reticula and Golgi bodies in cytoplasm. The nucleoplasm of secondary spermatocyte was more concentrated than that of the previous phase. Spermatids were more condensed in nucleus and cytoplasm, and show the long-spherical shape. In the cytoplasm of spermatid mitochondria located to lower portion of the nucleus and Golgi bodies located to upper portion, but proacrosomal granule is not appeared. The spermatozoon consist of the head and tail. No acrosome could be found in the head. The cytoplasmic collar of posterior part in sperm head contained mitochondria which surrounded axial filament. The well developed axonemal lateral fins were identified in sperm flagellum, and the axial filament of the flagellum consist of nine pairs of peripheral microtubules and one pair of central microtubules.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1412-1421
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• 2002

To examine the feasibility of using a sperm vector system for gene transfer, we have investigated the binding and the uptaking of foreign DNA into the sperm nucleus by PCR, in situ hybridization and LSC. We have also examined the transportation of exogenous DNA into oocytes by immunofluorescene via PCR. Sperm cells were incubated with DNA/liposome complexes (1:4 ratio) in fertilization medium with BSA or without BSA. In situ hybridization demonstrated that the transfection rate of sperm cells with and without BSA was 41 and 68% respectively, when the cells were treated with liposome/DNA complexes and 13% for DNA alone. LSC analysis showed that the binding of exogenous DNA was greatly reduced by DNase I treatment which digests DNA bound onto spermatozoa, suggesting that some of the DNA was internalized into the sperm membrane. To find out whether transfected DNA was internalized into sperm intracytomembrane, sperm DNA was amplified by inverse PCR. No PCR products were detected from sperm cells, indicating that the foreign DNA was simply bound onto the sperm membrane. To investigate transfer rates of exogenous DNA into oocytes via sperm cells, we used immunofluorescene method to follow the distribution of foreign DNA via spermatozoa: a few exogenous DNA was located in the cytoplasm of early embryos (13/60, 21.7% for DNA+/liposome+/BSA) and was not located in the pronucleus and/or nucleus. These results suggest that most of the transfected sperm cells could carry the foreign DNA into the egg by in vitro fertilization, but that the transferred DNA is degraded in the developing embryos without stable integration into the zygote genome. Therefore, we have directly injected with transfected sperm cell into oocyte cytoplasm and observed that some of the exogenous DNA was detected in preimplantation embryonic cytoplasm and expressed at preimplantation stages, suggesting that exogenous DNA in early zygote has their integrity. In this study, we have not identified a noble mechanism that interfering transportation of foreign DNA into zygote genome via spermatozoa. Our data, however, demonstrated that inverse PCR and immunofluorescene methods would be used as a new tool for follow-up of gene distribution in oocyte via sperm cells.

• Applied Microscopy
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• v.22 no.2
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• pp.97-117
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• 1992

In order to study process of spermiogenesis of the Korean greater horseshoe bat, Rhinolophus ferrumequinum korai, the cycle of seminiferous epithelium was examined by the light and electron microscope and the following results were obtained based on the epithelial cell differentiation. 1. Spermiogenesis occurred from early July to mid-Octber, and spermatogenic activity was vigorous from mid-August to late September. Spermatocytes including spermatogonia were found to be degenerated in only July. It is deduced that the degeneration serves as the mechanism to regulate effective use of energy to prepare for mating and hibernating periods, and regulation of breeding cycle. 2. Spermiogenesis of the Korean greater horseshoe bat was divided according to differentiation of the cell structure, into Golgi, cap, acrosome, maturation and spermiation phases; Golgi, cap and spermiation phases were further divided into two steps of early and late phase respectively, and acrosome phase into three steps of early, mid and late phases, and maturation phase has only one step. Hence, the spermiogenesis consists of ten phases. The first research was done in this article on the changes of chromatin with nucleus, the time of appearance and disappearance of chromatin granules, in case of Korean greater horseshoe bat (Rhinolophus ferrumequinum korai). Chromatin granule began to be condensed in late Golgi and the condensation proceeded to form an irregular mass of a electron-dense chromatin in a form of circular cylinder in the center of nucleus at the phase of maturation. Finally, the chromatin condensation proceeded and perfect nucleus of sperm with homogeneous density was formed when the sperm was separated from Sertoli cell. Therefore, appearance and disappearance of chromatin granules occurred in the period of time between late Golgi and maturation phase, The tail of sperm began to develop in early cap phase, Numerous lipid droplets were obseved in the cytoplasm of spermatids during the maturation phase, which seemed to be used as energy source necessary to make mature sperm during spermiogenesis.

• Applied Microscopy
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• v.32 no.4
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• pp.303-310
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• 2002

Spermatogenesis and sperm ultrastructure are investigated by means of light and transmission electron microscopy in the equilateral venus, Gomphina veneriformis which is dominant bivalve in the east coast of Korea. In the active spermatogenic season, testis consists of numerous spermatogenic follicles which is contains germ cells in the different developmental stage. The spermatogonia attached to spermatogenic follicle wall and has a large nucleus with electron-dense nucleolus. The spermatocytes are characterized by appearance of synaptonemal complex and well-developed Golgi complex. Nucleus of spermatid consists of numerous heterogeneous granules with high electron density. Karyoplasmic condensation, acrosome and flagellum formations are observed during spermiogenesis. Testicular matured sperms of sperm bundle consists of head, midpiece and tail. The head is about $8.5{\mu}m$ long and comprises a long nucleus and a bullet-like acrosome ($8.5{\mu}m$ in length). Acrosomal rod of microfilaments is observed in the lumen between nucleus and acrosome. The midpiece has four mitochondria. And tail has the typical '9+2' microtubule system.

• Journal of Aquaculture
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• v.10 no.4
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• pp.381-386
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• 1997

A series of experiments were conducted to compare the effects of various diluents in cold storage on the marbled sole, Limanda yokohamae sperm. Various diluents of glucose, L. yokohamae serum, marine fish Ringer's solution, sodium citrate and Ca2+ free artificial seawater (ASM) (tris-HCI, pH 7.4) containing 3 mM ethylene glycol-bis (2-aminoethyl ether) tetraacetic acid (EGTA) were used to store the sperm at 4℃. The storage effect was evaluated using motility index and survival rate of sperm. Glucose and sodium citrate were found to be better diluents which maintained high motility and survival rate of sperm for a storage period of 10 days. Some morphological changes of spermatozoa were observed during the cold storage with diluents. In particular, a detachment of the nuclear enveloped and of the plasma membrane from the nucleus in spermatozoa was observed. Morphological normality of the stored spermatozoa diluted with 0.3 M glucose was better than that of the stored spermatozoa undiluted or diluted with Ca2+ free ASW containing 3 mM EGTA.

• The Korean Journal of Malacology
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• v.27 no.2
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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.

• Development and Reproduction
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• v.16 no.1
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• pp.19-29
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• 2012

The ultrastructural characteristics of germ cell differentiations during spermatogenesis and mature sperm morphology in male $Atrina$ ($Servatrina$) $pectinata$ were evaluated via transmission electron microscopic observation. The accessory cells, which contained a large quantity of glycogen particles and lipid droplets in the cytoplasm, are assumed to be involved in nutrient supply for germ cell development. Morphologically, the sperm nucleus and acrosome of this species are ovoid and conical in shape, respectively. The acrosomal vesicle, which is formed by two kinds of electron-dense or lucent materials, appears from the base to the tip: a thick and slender elliptical line, which is composed of electron-dense opaque material, appears along the outer part (region) of the acrosomal vesicle from the base to the tip, whereas the inner part (region) of the acrosomal vesicle is composed of electron-lucent material in the acrosomal vesicle. Two special characteristics, which are found in the acrosomal vesicle of A. ($S$) $pectinata$ in Pinnidae (subclass Pteriomorphia), can be employed for phylogenetic and taxonomic analyses as a taxonomic key or a significant tool. The spermatozoa were approximately $45-50{\mu}m$ in length, including a sperm nucleus (about $1.43{\mu}m$ in length), an acrosome (about $0.51{\mu}m$ in length), and a tail flagellum (about $46-47{\mu}m$). The axoneme of the sperm tail evidences a 9+2 structure.

• Applied Microscopy
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• v.41 no.1
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• pp.31-35
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• 2011

The fine structures of the sperm morphology in the Myotis daubentonii ussuriensis were observed by transmission electron microscope. The results showed that the sperm head revealed bullet shaped, the width was showed a slender more than toward the posterior region to anterior region of nucleus. The sperm head was about $4.5{\mu}m$ in length, being about $2.0{\mu}m$ in width. The nuclear length was $4.3{\mu}m$, occupied most of the sperm head. The nucleus and acrosome were separated by the apical body. The neck region was composed the basal plate, capitulum and segmented columns. The segmented columns were about 12 to 14 in number and connected with the outer dense fibers of the middle piece. The mitochondria sheath were arranged like the thread of a screw, and the total number of mitochondrial gyres were 57. The satellite fibers were observed irregularly among the outer dense fibers in the middle piece. Except the middle piece they are not observed in the principal and end pieces of the tail. In general, the tail show axoneme composed of a 9+2 microtubular pattern, and microtubules of the end piece were arranged irregularly.

• Applied Microscopy
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• v.35 no.4
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• pp.42-54
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• 2005

This study described the spermatozoa of the discoglossidae Bombina orientalis using light, scanning and transmission electron microscopes. Sperm head possess a crescent or leaf shape, with a moderately flexible head, and with a sharp anterior and posterior tips. The nucleus are a thick cone shaped in the widest middle part of nucleus, and a slender anterior and posterior of nuclear tips. The chromatin is not completely compact, but irregularly imbricated such as roof. Some nuclear lacunae, irregular in shape, are scattered within the nucleus. No neck and middle piece were developed. The flagellum is composed of 9+2 axoneme, axial rod and undulating membrane. The mitochondria were distributed only in cytoplasmic membrane around the nucleus. In particular, the nuclear rod contains bundles of fibers, the rod penetrating from anterior portion to the middle of the nucleus, is extended roughly two-thirds of the nucleus such as eyelashes shaped.