• Asian-Australasian Journal of Animal Sciences
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• v.29 no.10
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• pp.1407-1415
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• 2016

Isolation and culture of spermatogonial stem cells (SSCs) are attractive for production of genetic modified offspring. In the present study, buffalo spermatogonial stem-like cells were isolated, cultured and expression pattern of different germ cell marker genes were determined. To recover spermatogonia, testes from age 3 to 7 months of buffalo were decapsulated, and seminiferous tubules were enzymatically dissociated. Two types of cells, immature sertoli cell and type A spermatogonia were observed in buffalo testes in this stage. Germ cell marker genes, OCT3/4 (Pou5f1), THY-1, c-kit, PGP9.5 (UCHL-1) and Dolichos biflorus agglutinin, were determined to be expressed both in mRNA and protein level by reverse transcription polymerase chain reaction and immunostaining in buffalo testes and buffalo spermatogonial stem-like cells, respectively. In the following, when the isolated buffalo buffalo spermatogonial stem-like cells were cultured in the medium supplemented 2.5% fetal bovine serum and 40 ng/mL glial cell-derived neurotrophic factor medium, SSCs proliferation efficiency and colony number were significantly improved than those of other groups (p<0.05). These findings may help in isolation and establishing long term in vitro culture system for buffalo spermatogonial stem-like cells, and accelerating the generation of genetic modified buffaloes.

• The Korean Journal of Cytopathology
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• v.8 no.2
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• pp.185-189
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• 1997

A sex cord tumor with annnular tubules is a relatively rare ovarian neoplasm. The cytologic findings from a fine needle aspiration biopsy of neck metastasis of a sex cord tumor with annnular tubules are described. The origin of the neck metastasis was the right ovary, and the tumor was diagnosed six years ago. The cytologic findings were characterized by tumor cells arranged in solid or follicular patterns. The tumor cells formed rosette-like or complex tubular structures with central rounded or coalesced hyaline materials. It was difficult to distinguish this tumor cytologically from granulosa cell tumor, thyroid follicular neoplasm, Sertoli-Leydig cell tumor, and Brenner tumor, but complex tubular structures were helpful in discriminating between these tumors.

• 대한생식의학회:학술대회논문집
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• 2006.06a
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• pp.156.2-156.2
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• 2006

• Development and Reproduction
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• v.26 no.2
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• pp.59-69
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• 2022

Many efforts have been made to study the expression of aquaporins (AQP) in the mammalian reproductive system, but there are not enough data available regarding their localized expression to fully understand their specific roles in male reproduction. The present study investigated the expression and localization patterns of different AQP subtypes in the adult mouse testes and testicular spermatozoa using an immunofluorescence assay. All the studied AQPs were expressed in the testes and revealed subtype-specific patterns in the intensity and localization depending on the cell types of the testes. AQP7 was the most abundant and intensive AQP subtype in the seminiferous tubules, expressing in Leydig cells and Sertoli cells as well as all stages of germ cells, especially the spermatids and testicular spermatozoa. The expression pattern of AQP3 was similar to that of AQP7, but with higher expression in the basal and lower adluminal compartments rather than the upper adluminalcompartment. AQP8 expression was limited to the spermatogonia and Leydig cells whereas AQP9 expression was exclusive to tails of the testicular spermatozoa and elongated spermatids. Taken together, the abundance and distribution of the AQPs across the different cell types in the testes indicating to their relavance in spermatogenesis, as well as in sperm maturation, transition, and function.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.225-230
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• 1999

In mouse, the heat shock protein 70-2 (hsp70-2) is found to have special function in spermatogenesis. Based on the observation, the hypothesis that human hspA2 (human gene; 98.2% amino acid homology with hsp70-2) might have important function in spermatogenesis in human testes was proposed. To test the hypothesis, we examined the expression of hspA2 in human tissues. Expression vector pDMC4 for expression of the human hspA2 protein using pTricHisB (invitrogen, USA) was constructed and the expressed hspA2 protein was cross-reacted with antiserum 2A raised against mouse hsp70-2 protein. Based on the cross-reactivity, we determined the expression level of hspA2 protein in human tissues by western blot analysis using the antiserum 2A. We demonstrated that antiserum 2A antibodies detected human hspA2 protein with specificity which was produced in the E.coli expression system. On Western blot analyses, significant hspA2 expression was observed in testes with normal spermatogenesis, whereas a low level of hspA2 was expressed in testis with Sertoli-cell only syndrome. Also, a small amount of hspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. These results demonstrate that the hspA2 protein is highly expressed in male specific germ cells, which in turn suggests that hspA2 protein might playa specific role during meiosis in human testes as suggested in the murine model. However, further studies should be attempted to determine the function of hspA2 protein in human spermatogenesis.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.3
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• pp.1351-1355
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• 2014

Background: Ovarian sex-cord stromal tumours (SCST) are rare, and relatively infrequent in children. These have to be distinguished from more common germ cell tumors in children and also from benign epithelial neoplasms. Objectives: The purpose of our study was to report the clinical and pathological findings in young patients with these tumours in our population. Material and Methods: The present observational cross-sectional study included all subjects <21 years of age diagnosed with ovarian SCST, in Aga Khan University Hospital Histopathology Laboratory, Karachi, Pakistan, from January 1992 till July 2013. Results: Of the total of 513 SCSTs presented during the study period, 39 fulfilled inclusion criteria and were assessed. The age range was 4-250 months. Most of the tumours presented at stage-1 and an abdominal mass was the most common presenting symptom, along with menstrual disturbance. The left side ovary was slightly more affected (53.5%). Of the total, 15 were juvenile granulosa cell tumours (JGCT), 11 sclerosing stromal tumours (SST), 10 of the fibrothecomas spectrum, 2 Sertoli leydig cell tumours (SLCT) and one a sex cord tumour with annular tubules (SCTAT). Detailed immunohistochemical analyses were performed in 33 cases. Recurrence/metastasis was noted in 4/21 cases with follow-up data. Conclusions: Ovarian sex cord stromal tumours are very rare in young age in our population, and usually present at an early stage. Most common among these are juvenile granulosa cell tumours, although surprisingly sclerosing stromal tumours were also common. Clinical symptoms due to hormone secretion in premenstrual girls and menstrual disturbance in menstruating girls are common presenting features.

• 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%.

• Applied Microscopy
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• v.24 no.3
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• pp.55-66
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• 1994

This study was carried out to investigate the histological changes of sperm cells in testis, obtained from 100 of 3-year-old male rainbow trout (Oncorhynchus mykiss) collected and analysed from March in 1992 to February in 1993. Especially, the ultrastructural changes of spermatogonia, primary and secondary spermatocytes, spermatids, and spermatozoa were examined to describe the reproductive cycles of this species. The results obtained in this study were as follows: The ultrastructures of the gonadotrophs largely parallel the cyclical changes in the testes. Each nest of cells belongs to one spermatogenetic stage, although nests at different stages can be found within the one lobule. At first keterochromatin is dispersed and then is condensed. In mature gamete, the nucleus is dense and homogeneous. The nuclear membrane appeared at the beginning of differentiation. In spermatogonia, Sertoli cells are located at the periphery of their cytoplasm. In the primary spermatocytes, the small mitochondria are abundant over the outer cytoplasm. During cell differentiation, the cytoplasm decreases and the nucleus increases. In spermatids, the protein masses moved towards the posterior part of the nucleus. In late spermatids, the two large mitochondria are located over the cytoplasm. In spermatozoa, two spheroidal mitochondria (about 145nm long) are situated in parallel between the nucleus and the axoneme. Spermatozoa mitochondria are assembled into an organized sheath surrounding the outer dense fibres and axoneme of the flagellar midpiece. The two centrioles are quite separate and the central pair and sheath complex of the flagellum is inserted into the base of the distal centriole.

• 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.

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.23-36
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• 1993

To investigate the cycle and relative frequences and the fine structure of seminiferous epithelia in mature Jindo dogs, histologic study was performed. The results obtained were summarized as follows; 1. Type A spermatogonia appeared approximately 1.6 times as many at stage II as compared to stage I while type In spermatogonia appeared small amount in stage III, IV and V. type B spermatogonia were found during the stage VI to VIII, though not detectable during stage I to V. The type B spermatogonia divided at stage VII to produce the preleptotene primary spermatocytes at stage VIII. The number of primary spermatocytes of the leptotene phase markedly increased during stage I to II, and the primary spermatocytes of the pachytene phase were shown the least in number at stage IV. The secondary spermatocytes could be seen only at stage IV. 2. The relative frequencies of each stage from stages I to VIII of the cycle of seminiferous epithelia were 31.6, 11.9, 10.0, 3.2, 8.2, 10.1, 11.7 and 13.2% respectively. 3. On electron microscopic observations, acrosomal vesicle of spermatids appeared larger though the bulk of germ cells were the morphologically same as those of the other animal species. Thread line structures light microscopically observed in the cytoplasm of Sertoli cell were the longitudinal orientation of mitochondria.