• Molecules and Cells
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• v.42 no.11
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• pp.794-803
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• 2019

Ultraviolet light (UV)-induced cellular response has been studied by numerous investigators for many years. Long noncoding RNAs (lncRNAs) are emerging as new regulators of diverse cellular process; however, little is known about the role of lncRNAs in the cellular response to UV treatment. Here, we demonstrate that levels of lncRNA-HOTTIP significantly increases after UV stimulation and regulates the UV-mediated cellular response to UV through the coordinate activation of its neighboring gene Hoxa13 in GC-1 cells (spermatogonia germ cell line). UV-induced, G2/M-phase arrest and early apoptosis can be regulated by lncRNA-HOTTIP and Hoxa13. Furthermore, lncRNA-HOTTIP can up-regulate ${\gamma}-H_2AX$ and p53 expression via Hoxa13 in UV-irradiated GC-1 cells. In addition, p53 has the ability to regulate the expression of both lncRNA-HOTTIP and Hoxa13 in vitro and in vivo. Our results provide new data regarding the role lncRNAs play in the UV response in spermatogenic cells.

• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.43-52
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• 1996

This study was carried out to establish the in vitro short-term culture system of developing male germ cells by purifing germ cells of various stages. The decapulated testicular cells were incubated with collagenase (lmg/ml) and try psin (2.5mg/ml) in HBSS. After separating male germ cell, the separated germ cells were stained with heamatoxylin/eosin and determined developing stages under light microscopy. The purity of pachtene spermatocytes a and round spermatid were 85%, respectively. Yield of total male germ cells was highly variable between individuals, with a mean value of 3.5 to 4.5 ${\times}$ 10$^7$ cells/testis. Viability of the cell was over 97% after separation. In DMEM medium, the optimal cell number for culture is approximately 1 x 10$^5$ cells/dish, but low cell den-sities than 1 ${\times}$ 10$^5$ cell/dish showed a decreased cell viability. Furthermore, about :36.8% of pac-hytene cells was successfully cultured for 6 days and some of cells were developed to secondary spermatids and round spermatids. Therefore, our data suggested that this culture conditions will be utilize as a feasible tools to produce tran-sgenic livestock using techniques such as intrac-ytoplasmic injection and cell fusion.

• The Korean Journal of Zoology
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• v.20 no.2
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• pp.67-76
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• 1977

The periodic changes of testis and \ulcorner션 epididymidis in Korean hibernating bats, the oriental discoloured bats (Vesportilio superans Thomas) and the Korean greater horseshoe bats (Rhinolophus ferrumequinum korak Kuroda) were studied in order to clarify the possibility of correlation between their histological findings and one of physiological facets, hibernation, and the results obtained were as follows: 1. The spermatogenic function of the oriental discoloured bats obtained in July and August was depressed although the spermatocytes showed a considerable cell proliferation. Few mature sperms were observed in the seminiferous tubules of the bat obtained in August. 2. The spermatogenic function of the Korean greater horeshoe bats obtained in September was not remalkable but a considerable number of sperms were stored in the excretory ducts which were characterized by existence of para-tubular spaces in the ductus epididymidis. 3. The spermatogenic epithelia of the Korean greater horseshoe bats obtained in December showed histologically atrophied figures. However, a vast number of sperm remained in extremely expaned luminae of the ductuc epididymidis which epithelial cells were maintained rectangular in shape. 4. These results suggest that there are periodic changes of the spermatogenic epithelia and the excretory ducts, and that those histological changes are closely related to their wintering.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.37-43
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• 1999

The image analyser was used for the measurement of the morphological changes of the spermatogenic cells and seminiferous tubules in the pheasant during the breeding and non breeding season. 1. The seminiferous tubules were enlarged 2.57 times during the breeding season than during the non breeding season. Only Sertoli cells and spermatogonia in the seminiferous tubules can be found during the non breeding season. 2. There is no significantly difference in the ratio of nucleus area against that of cell size in the spermatogonium between breeding and non breeding season. The ratio of area was 28.71% and 29.11%, respectively. However, the enlargement of spermatogonium was noticed during the non breeding season. 3. The highest value of the ratio of the nucleus area against that of cell size among the germ cells was measured 37.40% in the pachytene phase of the spermatocyte during the breeding season. 4. The ratio of nucleus area against that of cell size in the spermatid was 22.53%.

• 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.

• 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.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.45-51
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• 1999

The morphological study was carried out to classify the spermatogenic germ cells of the seminiferous tubule in Korean Native Pheasant during the breeding season. The results were as follows : 1. The spermatogonia can be classified into the three types of A, In(intermediate) and B. 2. The primary spermatocyte can be classified into the five types as preleptotene, leptotene, zygotene, pachytene and diakinesis. 3. The maturing processes of nucleus of spermatid can be divided into seven steps. The round shape of the spermatid was changed to the elongated form during the spermiogenesis. This observation may be useful to the study of the breeding cycles in the Korean Native Pheasant.

• 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.

• Journal of Embryo Transfer
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• v.28 no.1
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• pp.73-78
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• 2013

The present study examined the histological characteristics of adult testis in the long-beaked common dolphin (Delphinus capensis) from Korean waters and the localization of DEAD-box polypeptide 4 (DDX4; a germ cell marker) and vimentin (a Sertoli cell marker) expression in the dolphin testis compared with that in terrestrial mammals, including dogs and rats. The seminiferous tubules of dolphin testis have very small or completely closed lumens, and spermatogenic cells and Sertoli cells within the tubules cannot be differentiated. Immunohistochemical analysis showed that, in the dolphin testis, DDX4- and vimentin-positive cells were scattered extensively within the tubule, whereas in the dog and rat testis, DDX4 immunoreactivity was localized in spermatogenic cells of the adluminal compartment, and vimentin immunoreactivity was localized in Sertoli cells of the basal compartment in the seminiferous epithelium. These results suggest that the histological characteristics of the seminiferous tubules in the dolphin testis differ from those of terrestrial species.

• 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.