• Animal cells and systems
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• v.1 no.2
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• pp.371-379
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• 1997

To investigate the movement of sperm head and the role of sperm neck in forward sperm motility in the Korean striped field mouse, Apodemus agrarius coreae, the morphological characteristics of the cauda epididymal spermatozoa were examined by light microscopy and scanning and transmission electron microscopy. Spermatozoa of A. agrarius coreae were characterized by the conspicuous shape of the acrosome and the long tail compared with those of other rodents. Total length of the sperm was $133\mu{m}$. The sperm head had a curved falciform shape. The head was 8.0${\mu}$m in length, and about 4.0 ${\mu}$m in width. The shape of acrosome had an openerlike form. The sperm tail (125 ${\mu}$m) consisted of four major segments: neck (0.5 ${\mu}$m), middle piece (29.5 ${\mu}$m), and principal piece plus the end piece (95 ${\mu}$m). The outer dense fibers were arranged in a horseshoe fashion, and No. 1, 5, 6, and 9 of the outer dense fibers were larger than the others. The mitochondrial bundles of middle piece were composed of a pair of arms, which surrounded the axone of the middle piece by the 45 0 angled helical structure. The total number of mitochondrial gyres was 188. In particular, the microfilament structures existed in plasma membrane of the sperm, which was adjacent to the acrosomal region on the nuclear membrane. The segmented columns were surrounded by microfilament structures, and the microfilament bundles were adjacent to the outer membrane of the first mitochondria of middle piece. This study presents for the first time the existence of microfilament structures within the plasma membrane of sperm which is located from the adjacent acrosome region to the connecting piece in sperm neck of Korean striped field mouse, Apodemus agrarius coreae. The present result suggests that the constriction and extension of microfilament in sperm neck as well as the wave-movement of sperm tail may play a role in the movement of sperm head.

• Journal of Embryo Transfer
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• v.29 no.4
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• pp.351-359
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• 2014

Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.189-194
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• 1997

The ovaries of Korean native cows or heifers were obtained from a slaughter house and kept on 28~3O˚C and transported to laboratory within 2 hrs. The follicular oocytes were collected follicles. The oocytes were matured in vitro for 24 hrs. In TCM-199 supplemented with 35 $\pi$g /ml FSH, 10 $\pi$g /ml LH, 1 $\pi$g /ml estradiol-17 and granulosa cells at 39˚C under 5% $CO_2$ in air. The caudal epididymis of Korean native bulls were obtained from a slaughter house and transported to laboratory within 30 minutes. Swim-up of collected spermatozoa and freezing sperm was layered under 2ml fertilization B. 0. medium in two tissue culture tubes and held at a 45˚C angle for 0~2 hrs. They wrer fertilized in vitro by freezing sperm treated with heparin for 24 hrs, and then the zygotes were co-cultured in vitro with bovine oviductal epithelial cells for 7 to 9 days. The follicular oocytes recovered were classified into 41.7% as grade I, 51.5% as grade II and 6.8% as graed III. The number of oocytes recovered per ovary was averaged 8.3 and they were classifed into 2.3 as grade I, 2.5 as grade II and 2.3 as grade III. The cleavage rate of matured oocytes was significantly(P

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.256-256
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• 2004

The study was carried out to investigate the effects of preservation of ovaries and oocytes with and without cumulus cells and incubation time on zona penetration by canine spermatozoa. The objective of this study was to produce in vitro fertilized oocytes and solute canine sterile. (omitted)

• Development and Reproduction
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• v.25 no.1
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• pp.1-14
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• 2021

The Syrian (golden) hamsters are seasonal breeders whose reproductive functions are active in summer and inactive in winter. In experimental facility mimicking winter climate, short photoperiod (SP) induces gonadal regression. The blood-testis barrier (BTB) of the sexually involuted animals have been reported to be permeable, allowing developing germ cells to be engulfed or sloughed off the epithelium of the seminiferous tubules. The expressions of genes related to the tight junction composing of BTB were investigated in the reproductive active and inactive testes. Claudin-11, occludin, and junctional adhesion molecule (JAM) were definitely expressed in the active testes but not discernably detected in the inactive testes. And spermatozoa (sperm) were observed in the whole lengths of epididymides in the active testes. They were witnessed in only cauda region of the epididymides but not in caput and corpus regions in animals with the inactive testes. The results imply that the disorganization of BTB is associated with the testicular regression. The developing germ cells are swallowed into the Sertoli cells or travel into the lumen, as supported by the presence of the sperm delayed in the last region of the epididymis. These outcomes suggest that both apoptosis and desquamation are the processes that eliminate the germ cells during the regressing stage in the Syrian hamsters.

• Biomedical Science Letters
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• v.23 no.2
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• pp.128-132
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• 2017

Streptozotocin (STZ), bisphenol A (BPA), and diethylstilbestrol (DES) are known as endocrine disruptors, occurs oxidative stress in animal cells. Generally, oxidative stress induces reactive oxygen species (ROS) and lipid peroxidation of sperm and lead to decreased viability and fertility in pigs. Therefore, we investigated the influence of STZ, BPA and DES on ROS production and lipid peroxidation on boar sperm. Collected sperm were incubated with semen extender containing $10{\mu}M\;STZ$, $10{\mu}M\;BPA$ and $20{\mu}M\;DES$ for 3, 6 and 9 hours. Intracellular ROS and lipid peroxidation of sperm were analyzed by 2', 7'-dichlorofluorescein diacetate and malondialdehyde methods. The results show that, intracellular ROS was not significantly different among the all treatments, but lipid peroxidation was significantly increased in STZ group at 3 hour after incubation with boar sperm (P<0.05). These results suggest that STZ stimulates lipid peroxidation more than ROS production and may exert a negative effect on sperm fertility.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.3
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• pp.171-175
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• 2000

The ultrastructures of germinal cells of male marsh clam, Corbicujar lena were studied. The mature sperm was primitive type, consisting of head, middle piece and tail. The mature sperm was whip-shaped and its head was divided into two parts; the acrosomal part shaped long hollow cone about $3{\mu}m$ in length and the sperm nuclear part shaped a long stick about $9\;{\mu}m$ in length. The posterior part of the sperm nuclear projected to centriole, The middle piece of the sperm-nuclear had four mitochondria and two centrioles. The sperm tail part had the 9+2 microtubular arrangement known as a typical pattern, During spermiogenesis, chromatin within sperm nuclear became fiberic materials by condensation.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.333-335
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• 1999

This study was carried out to find out the effect on fertilizing capacity according to sperm concentration of liquid boar semen. Four different doses with various motile sperm cells of 3.0$\times$10$^{9}$ , 2.5$\times$10$^{9}$ , 2.0$\times$10$^{9}$ , and $1.5\times$10$^{9}$ per 80$m\ell$ plastic bottle were inseminated twice 12 h interval after standing estrus in 6,818 sows. Farrowing rate and total piglets per litter were 82.2% and 10.9, respectively, with no significant differences among the other treatments. The presumption of optimal concentration of motile sperm cells in the liquid boar semen was best at 2.0~2.3$\times$10$^{9}$ per dose.

• Applied Microscopy
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• v.30 no.1
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• pp.21-44
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• 2000

This study was carried out to investigate sperm storage, and the fate of spermatozoa in the female reproductive tract during hibernation in Korean greater horseshoe bat, Rhinolophus ferrumequinum korai. (1) Numerous sperm occurring in uterine lumen and glands were engulfed, and disappeared by the polymorphouclear leucocytes during the hibernation. (2) The stored sperm present in caudal isthmus of oviduct only, the heads of sperm toward the oviductal epithelial cells. Therefore, the projected sperm during the mating season are only alive in the caudal isthmus of oviduct in the long hibernation. The present result suggests that the caudal isthmus of oviduct may play an important role as the principal storage site in capacitation of sperm. (3) In March, the sperm do not occur in the caudal isthmus of oviduct. It suggests that the stored sperm in the caudal isthmus of oviduct should migrate to the ampulla of the site of fertilization to meet ovum in the period of ovulation. The results of this experiment consider that prolonged sperm storage, fate of sperm and sperm migration in the long hibernation have a kind of mechanism for the fertilization.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.1
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• pp.44-52
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• 2023

Objective: The DNA integrity of spermatozoa that attach to fallopian tube (FT) cells is higher than spermatozoa that do not attach. FT epithelial cells can distinguish normal and abnormal sperm chromatin. This study investigated the effects of sperm with a high-DNA fragmentation index (DFI) from men with unexplained repeated implantation failure (RIF) on the Toll-like receptor (TLR) signaling pathway in human FT cells in vitro. Methods: Ten men with a RIF history and high-DFI and 10 healthy donors with low-DFI comprised the high-DFI (>30%) and control (<30%) groups, respectively. After fresh semen preparation, sperm were co-cultured with a human FT epithelial cell line (OE-E6/E7) for 24 hours. RNA was extracted from the cell line and the human innate and adaptive immune responses were tested using an RT2 profiler polymerase chain reaction (PCR) array. Results: The PCR array data showed significantly higher TLR-1, TLR-2, TLR-3, TLR-6, interleukin 1α (IL-1α), IL-1β, IL-6, IL-12, interferon α (IFN-α), IFN-β, tumor necrosis factor α (TNF-α), CXCL8, GM-CSF, G-CSF, CD14, ELK1, IRAK1, IRAK2, IRAK4, IRF1, IRF3, LY96, MAP2K3, MAP2K4, MAP3K7, MAP4K4, MAPK8, MAPK8IP3, MYD88, NFKB1, NFKB2, REL, TIRAP, and TRAF6 expression in the high-DFI group than in the control group. These factors are all involved in the TLR-MyD88 signaling pathway. Conclusion: The MyD88-dependent pathway through TLR-1, TLR-2, and TLR-6 activation may be one of the main inflammatory pathways activated by high-DFI sperm from men with RIF. Following activation of this pathway, epithelial cells produce inflammatory cytokines, resulting in neutrophil infiltration, activation, phagocytosis, neutrophil extracellular trap formation, and apoptosis.