• Biomedical Science Letters
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• v.2 no.1
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• pp.57-69
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• 1996

To examine the process of spermiogenesis in the Korean red-backed vole, Clethrionomys rufocanus regulus, the seminiferous epithelium in the testis, was studied by trasmission electron microscopy, and the following results were obtained based on the morphological characteristics of cell differentiation. 1. According to the fine structural differentiation, spermiogenesis was divided into Golgi, cap, acrosome, maturation and spermiation phases. Besides, these phases were sub-divided into two steps : early and late phases respectively, Thus, the spermiogenesis of Clethrionomys rufocanus regulus was divided into a total of ten steps. 2. In the changes of the chromatin, the chromatin granules began to be condenced in the late Golgi phase, regularizated at maturation phases, and a perfect nucleus of sperm was formed at the spermiation phases. 3. The sperm head had the falciform, and the formative period of sperm tail began to be develop in the early Golgi phase and completed at the spermiation phases. 4. In the morphological features of spermiation phases, the spermatid of early spermiation phase was divided into three types : (1) A-type spermatid contained cytoplasmic droplets in the neck region and middle piece, and the mitochondria was irregular, and arranged around the axoneme. (2) B-type contained cytoplasmic droplet in the middle piece only, and the mitochondria are arranged the center of axoneme regularly, and (3) In the C-type spermatid, the arrangement of mitochondria was regular, and was contained cytoplasmic droplet in the neck region only. In the late spermiation phase, only the sperm head was surrounded by cytoplasm of Sertoli cell or the matured sperm just before the spermiation from the cytoplasm of Sertoli cell.

• Animal Bioscience
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• v.36 no.3
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• pp.385-403
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• 2023

Male fertility is affected by multiple endogenous stressors, including reactive oxygen species (ROS), which greatly deteriorate the fertility. However, physiological levels of ROS are required by sperm for the proper accomplishment of different cellular functions including proliferation, maturation, capacitation, acrosomal reaction, and fertilization. Excessive ROS production creates an imbalance between ROS production and neutralization resulting in oxidative stress (OS). OS causes male infertility by impairing sperm functions including reduced motility, deoxyribonucleic acid damage, morphological defects, and enhanced apoptosis. Several in-vivo and in-vitro studies have reported improvement in quality-related parameters of sperm following the use of different natural and synthetic antioxidants. In this review, we focus on the causes of OS, ROS production sources, mechanisms responsible for sperm damage, and the role of antioxidants in preserving sperm fertility.

• Journal of Embryo Transfer
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• v.28 no.2
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• pp.127-132
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• 2013

The objective of this study was to examine the effect of in vitro maturation (IVM) medium, cytochalasin B (CB) treatment during intracytoplasmic sperm injection (ICSI), and electric activation on in vitro development ICSI-derived embryos in pigs. Immature pig oocytes were matured in vitro in medium 199 (M199) or porcine zygote medium (PZM)-3 that were supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones for the first 22 h and then further cultured in hormone-free medium for an additional 21~22 h. ICSI embryos were produced by injecting single sperm directly into the cytoplasm of IVM oocytes. The oocytes matured in PZM-3 with 61.6 mM NaCl (low-NaCl PZM-3) tended to decrease (0.05

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.26-26
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• 2003

Hyaluronic acid (HA)-binding sites have been shown the diagnostic potential fur assessment of sperm maturity, which is related to male fertility. This study was designed to evaluate chromosomal patterns in porcine embryos produced by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with non- or HA-binding sperm (HABS). For binding of sperm with HA, sperm incubated in 10 ${mu}ell$ drop containing HA (0.8 mg/ml)-agarose (0.8%) mixture for 15 min. IVF and ICSI with non- or HA-bound sperm examined with matured oocytes at 44 hr after in vitro maturation. Embryos were cultured in 50 ${mu}ell$ of NCSU 23 containing 0.5% BSA for 5 days and then in 50 ${mu}ell$ of NCSU 23 containing 10% FBS for 2 days. For the evaluation of chromosomal aneuploidies, chromosome 1 sub-metacentric specific probe was used in sperm and embryos by fluorescence in situ hybridization (FISH). The frequency of aneuploidy sperm for chromosome 1 was 6.25%. The significant differences following IVF and ICSI with non- or HA-bound sperm were not observed in blastocyst formation rates (18.6, 23.5, and 23.8%) and cell number (61.8 $\pm$ 12.5, 55.5 $\pm$ 7.3, and 59.3 $\pm$ 9.6). Moreover, the percentage of diploidy in 4-cell stage embryos was 57.1% (IVF), 68.8% (ICSI), and 76.3% (ICSI-HABS). These results suggest that HA-binding sites may be a material for selection of normal sperm for ICSI. Therefore HA selection of normal sperm may be reduce the loss to embryonic mortality prior to embryo transfer in pig.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.3
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• pp.296-301
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• 2017

We conducted a study to 1) determine the indicators of gonadal maturity in male and female Korean walleye pollock Theragra chalcogramma for the purposes of artificial insemination; 2) establish the RNA/DNA ratio of mature eggs in this species; and 3) monitor the effect of refrigerated storage on egg viability and the motility of sperm collected from dead adult males. During the spawning season, the color of female gonads changed from orange to transparent, and that of male gonads changed from pale orange to milky white. The DNA content and RNA/DNA ratio of mature eggs were maintained without significant changes for approximately 6 h when eggs were preserved at $4^{\circ}C$. Sperm could be obtained from both milt and undiluted semen. Sperm obtained from milt ceased moving on the second day after isolation, while over 60% of sperm obtained from semen showed movement until the 13th day. Seven attempts were made to artificially inseminate mature eggs, of which two resulted in successful fertilization. The successful inseminations produced 94,000 and 5,000 fertilized eggs, respectively. This study shows that artificial insemination of walleye pollock is a viable strategy when natural propagation is not possible.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.1
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• pp.22-27
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• 2017

Objective: Azoospermia owing to testicular disorders is the most severe manifestation of male infertility. The main concern for patients with nonobstructive azoospermia (NOA) is the probability of successful sperm retrieval following testicular sperm extraction (TESE). Therefore, the goal of this study was to determine predictive factors correlated with sperm retrieval. Methods: We assessed the testicular histopathological patterns, the choice of TESE surgical procedure, hormone levels, and chromosomal abnormalities in patients with NOA (n=170). The histopathology specimens were analyzed based on the histopathological patterns of hypospermatogenesis, maturation arrest, and Sertoli cell-only syndrome. Results: The mean rate of sperm retrieval was 48.8%. The rate of sperm retrieval was significantly higher in the hypospermatogenesis group than in the other groups (p<0.001). There was a positive correlation between micro-TESE (vs. conventional TESE) and the sperm retrieval rate (odds ratio, 8.077; p<0.01). A logistic regression model demonstrated that high levels of follicle-stimulating hormone (FSH) and small testicular volume were significantly associated with lower chances of successful sperm retrieval. Conclusion: Some parameters, including testicular histopathology patterns, FSH levels, testicular volume, and method of TESE surgery, may be able to predict the chances of obtaining spermatozoa in patients with NOA. However, despite the efficiency of some predictive models, the hope of retrieving any functioning spermatozoa may be sufficient to disregard predictive factors of the success of intracytoplasmic sperm injection in these patients.

• Applied Microscopy
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• v.31 no.3
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• pp.245-256
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• 2001

The morphological characteristics of spermatogenic cells during the spermiogenesis of Paradoxornis webbiana were studied by transmission electron microscope. Spermiogenesis of P. webbiana was divided into ten phase. The chromatin granules became fibrous granules at the Golgi phase, gradually condensed at the cap phases, condensed as a stick at the acrosomal phase, and finally, a perfect nucleus was formed at the maturation phase. The formation of sperm tail began at the early Golgi phase, and completed at the late maturation phase. In particular, the dense materials existed in the sperm neck, which is wedged between the tip of segmented columns and the first mitochondria of the middle piece. The axone in the neck were surrounded by the dense materials. The axonema in spermatozoon contains a 9+2 arrangement of microtubules: 9 doublets, and 2 central single microtubules. Mitochondrial bundles of middle piece were composed of a pair of arms, which surrounded the axone of the middle piece by the $15^{\circ}$ angled-helical structure. The outer membrane of mitochondria were surrounded by microtubules in plasma membrane of the sperm. The undulating membrane had a helical structure, and the sperm plasma membrane was surrounded by undulating membrane.

• Korean Journal of Animal Reproduction
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• v.27 no.4
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• pp.275-280
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• 2003

This study was carried out to investigate the effect of cysteamine addition during in vitro maturation, fertilization and culture of porcine oocytes. Oocytes were matured for the first 22 h in mTCM -199 media supplemented with or without 150 $\mu$M cysteamine. They then were matured for an additional 22 h in mTCM-199 media without hormones supplemented with or without 150 $\mu$M cysteamine. When cumulus-oocyte complexes (COCs) were matured in the mTCM-199 media supplemented with cysteamine, the rates of GVBD and maturation (metaphase II) were enhanced as compared to the media without the addition of cysteamine. Also, when COCs were matured in the mTCM-199 media supplemented with cysteamine, the rates of sperm penetration, male pronucleus formation, cleavage and blastocyst formation after in vitro fertilization were enhanced as compared to the media without the addition of cysteamine. In conclusion, it was suggested that oocytes matured for the first 22 h in mTCM-199 media supplemented with 150 $\mu$M cysteamine increased the rates of metaphase II, sperm penetration, male pronucleus and blastocyst formation were higher as compared to the media without addition of cysteamine.

• Applied Microscopy
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• v.31 no.2
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• pp.129-141
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• 2001

To investigate the spermiogenesis of the Saghalien Pygmy shrew (Sorex minutus gracillimus), the testis obtained from mature male shrew was studied by electron microscopy, and the following results obtained based on the morphological characteristics of cell differentiation of the seminiferous epithelium in the testis. According to the fine structural differentiation, spermiogenesis of S. minutus gracillimks was divided into Golgi, cap, acrosome, maturation and spermiation phases. Beside, the Golgi and cap phases were subdivided into three steps of early, middle and late phase respectively, and acrosome phase into two steps of early and late phase , and maturation and spermiation phases has only one step respectively. Thus, the spermiogenesis of S. minutus gracillimus was divided into a total of ten steps. The chromatin granules begin to be condensed in the acrosome phase, and a perfect nucleus of sperm was formed at the spermiation phase. Mancette were appeared from the late acrosome phase to the maturation phase. The formation of sperm tail began to develop in the late Golgi phase, and completed at the spermiation phase. Multivesicular bodies were appeared from the Golgi phase to the maturation phase, recognized with pale, pale and moderate, and dense at Golgi, cap and acrosomal and matulation phases respectively.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.4
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• pp.352-361
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• 2021

Objective: The study assessed the developmental potential of germinal vesicle (GV) oocytes subjected to in vitro maturation (IVM) after prematuration culture with cilostamide (a phosphodiesterase-3 inhibitor) and the impact of cilostamide exposure on the morphology of meiosis II (MII) oocytes and subsequent embryo quality. Methods: In total, 994 oocytes were collected from 63 patients. Among 307 GV oocytes, 140 oocytes were selected for the experimental group and 130 oocytes for the control group. The denuded GV-stage oocytes were cultured for 6 hours with cilostamide in the experimental group and without cilostamide in the control group. After 6 hours, the oocytes in the experimental group were washed and transferred to fresh IVM medium. The maturational status of the oocytes in both groups was examined at 26, 36, and 48 hours. Fertilization was assessed at 18 hours post-intracytoplasmic sperm injection. Embryo quality was assessed on days 3 and 5. Results: In total, 92.1% of the oocytes remained in the GV stage, while 6.4% converted to the MI stage (p<0.01) after cilostamide exposure. In both groups, more MII oocytes were observed at 36 hours (25.8% vs. 21.5%) than at 26 hours (10.8% vs. 14.6%) and 48 hours (13% vs. 7.9%) (p>0.05). With the advent of cilostamide, blastocyst quality was better in the experimental group than in the control group (p<0.05). Conclusion: Cilostamide effectively blocked nuclear maturation and promoted cytoplasmic growth. Prematuration culture with cilostamide enabled synchronization between cytoplasmic and nuclear maturity, resulting in better blastocyst outcomes.