• Molecules and Cells
• /
• 제42권11호
• /
• pp.794-803
• /
• 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.

• 한국가축번식학회지
• /
• 제20권1호
• /
• pp.43-52
• /
• 1996

본 연구는 생쥐의 고게정관내에 존재하는 분화단계의 정자세포를 발생단계별로 분리하여, 체외에서 단기간 배양체계를 확립하기 위하여 실시하였다. 8주령 이상된 생쥐의 정소로부터 정소막을 제거시킨 후, collagenase (1mg/ml), trypsin(2.5mg/ml)를 처리하여 곡세정관을 간질세포와 분리하여 배양액에 부유시켰다. 부유세포는 Celcep장치를 이용하여 세포크기와 밀도 차이에 의해 분화 단계별로 분리하였다. 회수된 세포의 균질성은 haematoxylin/eosin 또는 orcein으로 염색한 후, 광학현미경하에서 확인한 결과 약 85% pachytene spermatocyte와 round spermatid을 성공적으로 분리해냈다. 따라서 sedimentation velocity에 의해서 생쥐의 spermatogenic cell의 발달단계별 분리가 가능함을 알 수 있었다. 이러한 방법으로 분리된 pachytene spermatogenic cell들은 DMEM 배양액에서 6일 이상 배양한 결과 약 36%의 생존율을 보였다. 따라서, 분화단계별 정자 세포의 분리 및 배양체계의 확립은 웅성생식세포의 발생과정에 따른 생리 또는 분자생물학적 현상을 규명함은 물론 세포융합기술의 이용에 의한 형질전환동물 생산에의 응용을 통해 가축에 있어서의 형질전환 생산효율의 개선에 기여할 수 있으리라 사료된다.

• 한국동물학회지
• /
• 제20권2호
• /
• pp.67-76
• /
• 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.

• 한국가축번식학회지
• /
• 제23권1호
• /
• pp.37-43
• /
• 1999

계절번식 조류인 꿩을 대상으로 번식기와 비번식기에 따른 세정관과 각 정자형성세포들의 직경, 길이와 면적 등의 변화상을 영상분석기를 이용하여 계측하였다. 그 결과는 다음과 같다. 1. 비번식기 곡세정관은 번식기보다도 약 2.56 배(38.94%) 위축되었다. 비번식기 세정관내에는 세르툴리세포와 정조세포만이 출현하였다. 2. 번식기 정조세포 핵과 세포질의 면적비는 28.71%이었으며, 비번식기의 면적비는 29.11%로 유의성은 없었다. 하지만 비번식기의 정조세포핵과 세포질 면적, 직경 및 둘레는 번식기보다는 증가했다. 3. 번식기 정모세포의 발달과정 중에서 태사기 상태에서 세포면적에 대한 핵의 면적비는 37.40%로 각 정자형성세포들 중에서 가장 높았다. 4. 원형에 가까운 정자세포의 세포면적에 대한 핵 면적비는 22.53%이었다.

• 대한방사선기술학회지:방사선기술과학
• /
• 제13권2호
• /
• pp.51-51
• /
• 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.

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

• 한국가축번식학회지
• /
• 제23권1호
• /
• pp.45-51
• /
• 1999

본 연구는 성성숙한 번식기의 한국산 숫컷 꿩을 대상으로 번식연구의 기초자료를 얻기 위해 곡세정관의 정자형성세포를 분류하였다. 그 결과는 다음과 같다. 1. 정조세포는 A형, In형 및 B형의 3 가지 정조세포로 분류되었다. 2. 제 Ⅰ정모세포는 전세사기, 세사기, 접합기, 태사기 및 이동기의 5가지 형 제 1 정모세포로 분류되었다. 3. 정자세포는 핵의 변화에 따라서 둥근 정자세포에서 장축성장한 정자세포로 성장하는 과정을 7 단계로 분류되었다.

• Applied Microscopy
• /
• 제28권2호
• /
• pp.193-205
• /
• 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.

• 한국수정란이식학회지
• /
• 제28권1호
• /
• pp.73-78
• /
• 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.

• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
• /
• pp.96-96
• /
• 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.