• Advances in Traditional Medicine
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• v.8 no.2
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• pp.111-124
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• 2008

In the present study, an attempt has been made to assess whether the effect of benzene extract of Ocimum sanctum leaves on the ultrastructural changes in the epithelial cells of the cauda epididymis, its subsequent recovery in the seminiferous epithelium and fertility of male albino rats. Wistar strain male albino rats were orally administered benzene extract of 250 mg/kg body weight of O. sanctum leaves followed by subsequent recovery maintaining suitable controls for 48 days. Results indicate decrease in the weights of testis, epididymis and seminal vesicles. Other accessory organs were not affected. Total count, cell and nuclei diameters of germ cells and Leydig cells were reduced. Cauda epididymis exhibited significant reduction in epithelial height and nuclei diameter of epithelial cells. Cells showed vacuolization with exhibit of signs of degeneration. Ultra study revealed that, in general, the cauda epididymis was affected and in particular, the principal, clear and basal cells were highly disturbed. Further, there was decrease in the size of lipid droplets, mitochondria, Golgi complex, endoplasmic reticulum and accumulation of lysosomal bodies. Fertility performance test showed no implantation in female rats mated with O. sanctum treated rats. Moreover, their recovery after withdrawal of treatment was observed suggesting that the effect of the treatment is transient and reversible. A recovery period resulted in normal spermatogenesis and fertility, suggesting reversible antispermatogenic and antifertility effects of the plant.

• Proceedings of the Korean Aquaculture Society Conference
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• 2004.05a
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• pp.495-497
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• 2004

• Toxicological Research
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• v.19 no.2
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• pp.153-159
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• 2003

Localization of mercury compounds was investigated in selective regions of the male reproductive tract using autometallography. The results demonstrated that mercury was observed in Sertoli and Leydig cells in testis, but not in the epithelial cells of rete testis and germ cells. In the efferent ductule, mercury compounds were observed in the cytoplasmic compartments of epithelial cells in the proximal and common regions, while they were observed in the supranuclear cytoplasmic compartments in the conus region. In the epididymis, the compounds were observed in the cytoplasmic compariments of narrow and basal cells, but not in the principal cells of the initial segment. In contrast, the compounds were evenly detected in the cytoplasmic compartments of principal cells in the caput. In the corpus and caudal epididymis, the compounds were observed in the basal region of principal cells. The data shows that mercury is differentially accumulated in the male reproductive tract in a region-specific manner.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.213-222
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• 2023

Cryopreservation is an option for the preservation of pre- or post-pubertal female or male fertility. This technique not only is beneficial for human clinical applications, but also plays a crucial role in the breeding of livestock and endangered species. Unfortunately, frozen germ cells, including oocytes, sperm, embryos, and spermatogonial stem cells, are subject to cryoinjury. As a result, various cryoprotective agents and freezing techniques have been developed to mitigate this damage. Despite extensive research aimed at reducing apoptotic cell death during freezing, a low survival rate and impaired cell function are still observed after freeze-thawing. In recent decades, several cell death pathways other than apoptosis have been identified. However, the relationship between these pathways and cryoinjury is not yet fully understood, although necroptosis and autophagy appear to be linked to cryoinjury. Therefore, gaining a deeper understanding of the molecular mechanisms of cryoinjury could aid in the development of new strategies to enhance the effectiveness of the freezing of reproductive tissues. In this review, we focus on the pathways through which cryoinjury leads to cell death and propose novel approaches to enhance freezing efficacy based on signaling molecules.

• Animal cells and systems
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• v.10 no.4
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• pp.227-231
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• 2006

Micro-deletions at specific loci of the Y chromosome have been observed frequently in male infertility patients, suggesting that genes in these regions are involved in male germ cell development. DAZ is a representative male infertility gene at the AZFc locus of the Y chromosome. Since DAZ contains an RNA binding motif along with so-called a DAZ domain, it was proposed to participate in RNA metabolism during spermatogenesis. A mouse gene homologous to the human DAZ gene has been cloned and named Dazl (DAZlike). Dazl is autosomal and expressed in the testis and also at a low level in the ovary. Male mice homozygous for the Dazl null allele have small testes with a few spermatogonia and almost complete absence of germ cells beyond the spermatogonial stage, suggesting the requirement of Dazl for entry or progression through meiosis. However, its exact cellular functions have not been understood yet. In order to investigate cellular functions of Dazl, we decided to isolate candidate interacting protein genes of the mouse Dazl, using yeast two-hybrid screening. A number of candidate Dazlinteracting proteins have been isolated, such as Bprp, Acf, Hgs, Murr1, Nbak3 and Ranbp9, but dynein light chain 1 (Dlc1) was most predominant. A strong interaction of Dazl with Dlc1 suggests that Dazl might function as an mRNA adaptor to the dynein motor complex.

• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.65-70
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• 2004

In mammals, male and female germline stem cells are derived from primodial germ cells. Despite many efforts to identify stem cells from gonads, there has been little successe to identify germline stem cells yet. In this study, we isolate and characterized porcine germline stem cells using only stem cell markers that are prevalently expressed in various tissues. Gonadal cells derived from both male and female formed colonies and showed AP activities and different lectin binding properties. Pluripotency of germline stem cells was also identified by positive signals against putative stem cells markers such as SSEA-1 and SSEA-3. In addition, nestin was also found in primary gonad cells that have a similar morphology to the AP-positive cells. The nestin expression suggests that the germline stem cells may have similar expression of the prevalent stem cell markers found in other tissues. The demonstration of nestin expression together with pluripotent cell markers calls further investigation of the possible differentiation of nestin-positive cells into neurons.

• The Korean Journal of Malacology
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• v.27 no.4
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• pp.377-386
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• 2011

The ultrastructures of germ cells during spermatogenesis and sperm morphology in male Mya arenaria oonogai, which was collected on the coastal waters of Samcheonpo, south coast of Korea, were investigated by transmission electron microscopic observations. In the early stage of the spermatid during spermiogenesis, a few granules and a proacrosomal granule, which is formed by the Golgi complex, appear on the spermatid nucleus, and then it becomes a proacrosomal vesicle. Consequently, it becomes an acrosome by way of the process of acrosome formation. The morphologies of the sperm nucleus type and the acrosome of this species have a curved cylindrical type and cone shape, respectively. The spermatozoon is approximately $48-50{\mu}m$ in length including a curved cylinderical sperm nucleus (about $2.65{\mu}m$ long), an acrosome (about $0.64{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$ long). As some ultrastructural characteristics of the acrosomal vesicle, the peripheral parts of two basal rings show electron opaque part (region), while the apex part of the acrosome shows electron lucent part (region). These charateristics of the sperm belong to the family Myidae or some species of Veneridae in the subclass Heterodonta, unlike 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 acrosome structures. Exceptionally, In particular, a cylinder-like nucleus of the sperm is curved (the angle of the nucleus is about $20^{\circ}$), as seen in some species of Veneridae (range from $0^{\circ}-80^{\circ}$). The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appeared in most species except for a few species in Veneridae in the subclass Heterodonta. Cross-sectioned axoneme of the sperm tail flagellum shows a 9+2 structure: the axoneme of the sperm tail flagellum consists of nine pairs of peripheral microtubules at the periphery and a pair of central doublets at the center.

• The Korean Journal of Malacology
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• v.26 no.1
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• pp.33-43
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• 2010

The ultrastructure of germ cells during spermatogenesis and some characteristics of sperm morphology in male Mytilus coruscus, which was collected on the coastal waters of Gyeokpo in western Korea, were investigated by transmission electron microscope observations. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves in that it contains a short midpiece with five mitochondria surrounding the centrioles. The morphologies of the sperm nucleus type and the acrosome shape of this species have an oval and modified cone shape, respectively. In particular, the axial rod is observed between the nucleus and acrosome of the sperm. The spermatozoon is approximately $45-50{\mu}m$ in length including a sperm nucleus (about $1.46{\mu}m$ in length), an acrosome (about $3.94{\mu}m$ in length) and tail flagellum (approximately $40-45{\mu}m$). 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. Some special charateristics of sperm morphology of this species in the genus Mytilus are (1) acrosomal morphology, (2) the number of mitochondria in the midpiece of the sperm, and (3) the existence of a satellite. The axial rod appears in the acrosome and sperm nucleus as one of the characteristics seen in several species of the subclass Pteriomorphia, unlikely the subclass Heterodonta containing axial filament instead of the axial rod. The number of mitochondria in the midpiece of the sperm of this species in the family Mytilidae are five, as one of common characteristics appeared in most species in the family Mytilidae. Most of Mytilus species contain a satellite body which is attached to the proximal centriole in the middle piece of the sperm, as one of common characteristics of sperm morphology in the family Mytilidae.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.235-239
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• 2005

This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.

• Animal Bioscience
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• v.36 no.6
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• pp.973-979
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• 2023

Objective: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system, which is the most efficient and reliable tool for precisely targeted modification of the genome of living cells, has generated considerable excitement for industrial applications as well as scientific research. In this study, we developed a gene-editing and detection system for chick embryo sexing during the embryonic stage. Methods: By combining the CRISPR/Cas9 technical platform and germ cell-mediated germline transmission, we not only generated Z chromosome-targeted knockin chickens but also developed a detection system for fluorescence-positive male chicks in the embryonic stage. Results: We targeted a green fluorescent protein (GFP) transgene into a specific locus on the Z chromosome of chicken primordial germ cells (PGCs), resulting in the production of Z^GFP-knockin chickens. By mating Z^GFP-knockin females (Z^GFP/W) with wild males (Z/Z) and using a GFP detection system, we could identify chick sex, as the GFP transgene was expressed on the Z chromosome only in male offspring (Z^GFP/Z) even before hatching. Conclusion: Our results demonstrate that the CRISPR/Cas9 technical platform with chicken PGCs facilitates the production of specific genome-edited chickens for basic research as well as practical applications.