• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.97-97
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• 2002

Achieving of in vitro development for mammalian premature spermatogenic cells are very difficult. In-vitro culture of spermatogenic cells were then initiated in an effort to try to study in vivo spermatogenesis and to understand its molecular events. Recently, the morphogenetic changes of spermatocytes or spermatid by in-vitro culture system were achieved. (omitted)

• Development and Reproduction
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• v.20 no.3
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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.

• Applied Microscopy
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• v.28 no.2
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• pp.193-205
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• 1998

The Urechis unicinctus sperm and spermatogenic cells prepared from the testis are investigated to identify $\alpha-tubulin$ of axoneme microtubules using mouse monoclonal $anti-\alpha-tubulin$ as the first Ab and Gold(10nm) conjugated goat anti-mouse IgG as the Ab marker. The Ag-Ab reaction analyzed excellently the localization of $\alpha-tubulin$ and the gold particles incorporated with the proximal and distal centrioles, manchette microtubules, and flagellum. The gold particles can be also observed in the spermatogenic cells while the cells are still in sperm ball which is composed of a somatic cell and spermatogenic cells. The sperm ball is the functional unit of sperm production in U unicinctus testis. The spermatids are developed from the spermatogenic cells in the sperm ball and released into the testis cavity through a cortical cytoplasmic opening. The spermatid architectures are similar with the mature sperm of the testis cavity in aspects of shape of discoid acrosome, degree of nuclear condensation and ring type of mitochondrion. However, the distal centriole connecting with the flagella can be observed from the mature sperm while the both proximal and distal centrioles reveal only in the spermatids. The proximal centriole is directly connected with nuclear outer membrane during the stage of nuclear condensation and oriented perpendicularly to the distal centriole whose axis coinciding with the longitudinal axis of the spermatozoon. There are indications that the distal centriole is intimately associated with the polymerization of the flagellum. The manchette microtubules appear during spermatid development but the mature sperm have round head and no conspicuous middle piece.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.96-96
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• 2003

Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

• 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.

• Development and Reproduction
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• v.17 no.2
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• pp.87-97
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• 2013

The seminiferous epithelium cycle and developmental stages of spermatids in Clethrionomys rufocanus were observed under a light microscope. The seminiferous epithelium cycle was divided into 8 stages. Type Ad spermatogonia appeared through all stages. Type Ap, In, and B spermatogonia appeared in stages I, II, III, and IV. In the first meiosis prophase, the leptotene spermatocytes appeared from stage V, the zygotene spermatocytes in stages I, VI, VII, VIII, the pachytene spermatocytes from stages II to VI, the diplotene spermatocytes in stage VII. The meiotic figures and interkinesis spermatocytes were observed in stage VIII. Developing spermatids were subdivided into 10 steps, based on the morphological characteristics such as the acrosome formation changes in spermatozoa, nucleus, cytoplasm, and spermiation changes. The C. rufocanus spermatocytogenesis and spermiogenesis results displayed similar results with Apodemus agrarius coreae and A. speciosus peninsulae. Considering all the results, the spermatogenesis may be useful information to analyze the differentiation of spermatogenic cells and the breeding season.

• Journal of radiological science and technology
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• v.13 no.2
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• pp.51-51
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• 1990

This study was undertaken to observe the effects of $^{60}Co\;{\gamma}-irradiation$ on the cell of spermatogenic epithelium in the testis of the chicken. 16-week-old chicken were provided as an experimental group and compared with control group. The experimental group was divided into a single irradiation (800, 1000, 1200 rads) and into three partial irradiation group (800/3, 1000/3, 1200/3 rads). The morphological changes of epithelial cell of the testis were observed by means of hematoxyline and eosin stain. Microstructure of spermatocyte and sperm was observed by means of semithin section of electron microscopic specimen. The results obstained are summerized as follows. 1. Spermatogonia and sertoli cells were found to be isolated from the basal membrane of seminiferous tubules as dose of $^{60}Co\;{\gamma}-irradiation$ was increased. 2. Spermatocytes of pachytene stage were seperated from the cytotplasmic process of sertoil cell in case of 1000 rads of $^{60}Co\;{\gamma}-irradiation$. 3. Normal arrangement of the cell of spermatogenic epithelium was found in control group and only the partial irradiation group of 800 rads. Vaculation in the seminiferous was pronounced in case of a single irradiation group of 800 rads, but the irradiation group of 1000 rads and 1200 rads were found to be damaged severely in both a single and a partial dose.

• Fisheries and Aquatic Sciences
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• v.10 no.3
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• pp.143-149
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• 2007

This study describes spermatogenesis and sperm ultrastructure of the granular ark, Tegillarca granosa using light and electron microscopy. In the active spermatogenic season, the testis comprises many spermatogenic follicles that contain germ cells in different developmental stages. Primary spermatocytes in the pachytene stage are characterized by synaptonemal complexes. The early spermatids are characterized by the appearance of several Golgi bodies, increased karyoplasmic electron density, and tubular mitochondria. The mass of proacrosomal granules consists of numerous heterogeneous granules with high electron density that are about 20 nm in diameter. From the midstage of spermiogenesis, the well-developed mitochondria in the cytoplasm aggregate posterior to the nucleus and surround the proximal and distal centrioles. The proacrosomal granules condense and form a single acrosome with a thin envelope. During late spermiogenesis, the acrosome begins to elongate becoming conical. The sperm is approximately $35.0{\mu}m$ long and consists of a head, midpiece, and tail. The head comprises a round nucleus and a conical acrosome. A micro fibrous axial rod is observed between the nucleus and acrosome. The midpiece has a calyx-like structure with five mitochondria, and the tail, which has the typical "9+2" microtubular system, originates from the distal centriole.

• Toxicological Research
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• v.19 no.2
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• pp.83-90
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• 2003

Since histopathological examination was known to be the most sensitive evaluation for testicular toxicity, regulatory authorities have been published the guidelines on practical testicular assay approach. Those guidelines specified details of evaluation including fixation, embedding, stain-ing, histological examination and also seminiferous tubular staging methods. However, there have been confusing understanding among toxicologists and even pathologists on staging theory and its application on industrial testicular toxicity. Guidelines did not intend to conduct quantitative assay with staging but recommended the use of knowledge of staging. To count each tubular stage with statistical analysis is known to be time consuming and labor burdening work but the significance of toxicity has little value. It also has been pointed out that the application of staging theory for longer-term toxicity considered to be lacking of rationale. It could be recommended that qualitative assay with aware-ness of germ cell loss is more efficient method rather than quantitative counting of each tubular stage. Therefore it would be required that comprehensive understanding of testicular toxicity evaluation and the use of testicular staging method.

• The Korean Journal of Malacology
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• v.29 no.1
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• pp.65-76
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• 2013

We investigated the gametogenic cycle and spawning seasons of the male Chlamys (Azumapecten) farreri nipponensis by qualitative and quantitative analyses, and also the size at 50% of group sexual maturity was calculated by the data of first sexual maturity. In this study, the male gametogenic cycle of this species by qualitative analysis was divided into five successive stages: early active stage (January to March), late active stage (March to April), ripe stage (April to August), partially spawned stage (July to September), and spent/inactive stage (August to January). The male gametogenic cycle showed similar patterns with monthly changes in the gonadosomatic index and condition index. Particularly, spawning in male scallop occurred once a year from July to September, unlike the spawning period of this species (from June to August) reported by the previous researchers. In quantitative statistical analysis using an image analyzer system, the patterns of monthly changes in the percent (%) of the areas occupied by spermatogenic stages to the testis areas in males showed a maximum in June, and then sharply dropped from July to September, 2006. From these data, it is apparent that the spawning season of C. (A.) farreri nipponensis occurred once per year from July to early September, indicating a unimodal gametogenic cycle during the year. Shell heights at 50% of group sexual maturity (RM50) fitted to an exponential equation were estimated to be 49.90 mm in males (considered to be one year old), and it was 100% for male scallops over 61.0 mm (considered to be two years old).