• 한국수정란이식학회지
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• 제14권2호
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• pp.131-137
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• 1999

This experiment was carried out to investigate the effects of osmolarity of thawing diluents, seminal plasma added in thawing diluents on the sperm viability and the effects of thawing temperature, the temparature of the thawing diluents on the sperm viability and acrosomal morphology of boar spermatozoa by the straw method. The result obtained were summarized as follows: 1. The sperm viablilty after thawing of the frozen semen was shown greater in the high osmolarity(392~492mOsm) than low osmolarity(300mOsm) in thawing diluent. The added levels of seminal plasma in thawing diluent did not affect the viability of frozen-thawed boar semen. 2. In terms of thawing temperature, the sperm viability was shown higher in the frozen semen thawed at 5$0^{\circ}C$ for one min. (p<0.01) than those thawed at 2$0^{\circ}C$ or 37$^{\circ}C$ for one min. The sperm viability was not significant at the diluent temparature of 2$0^{\circ}C$or 37$^{\circ}C$ after thawing: but the sperm viability was higher in thawing diluent at 2$0^{\circ}C$ than in that at 37$^{\circ}C$. However, the effects of thawing temperature and diluent solution on normal acrosomal rate were not significant. 3. Cleavage rates of oocytes fertilized with frozen semen were 46.4% and 43.3%, respectively, which were thawed at 5$0^{\circ}C$ for one min. and then diluted in mBTS medium at 2$0^{\circ}C$or 37$^{\circ}C$. To sum up, the sperm viability was shown greater at the high of thawing diluents of frozen boar semen. In terms of thawing conditions, the sperm viability was shown greater, when semen was thawed at a high temperature for a short time and then diluted at the same temperature as that in the straw.

• 한국패류학회지
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• 제28권2호
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• pp.165-174
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• 2012

The morphology and taxonomic values of the sperm in male Chlamys (Swiftopecten) swiftii were investigated by transmission electron microscope observations. The morphologies and ultrastructures of the sperm nucleus and the acrosome of this species are the vase type and long cone shape, respectively. Spermatozoa are approximately $45-50{\mu}m$ long including a sperm nucleus (approximately $2.60{\mu}m$ long), an acrosome (about $0.63{\mu}m$ long), and a tail flagellum (approximately $44-47{\mu}m$ in long). The axoneme of the sperm tail shows a 9+2 structure. In this study, the right and left basal rings in the acrosomal vesicle of this species show electron opaque part (region), and also the anterior apex part of the acrosomal vesicle shows electron opaque part (region). These characteristics of the acrosomal vesicle were found in Pectinidae and other several families in subclass Pteriomorphia. The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appear in most species in Pectinidae in subclass Pteriomorphia. In addition, the satellite fibres are found near the distal centriole of this species, as have been reported in other species of Pectinidae in subclass Pteriomorphia. Accordingly, structutral characteristics which are found in the acrosomal vesicle, four mitochondria in the sperm midpiece and the appearance of the satellite fibers near the distal centriole of C. (S.) swiftii in Pectinidae (subclass Pteriomorphia), can be employed for phylogenetic and taxonomic analyses as taxonomic key or a significant tool.

• Clinical and Experimental Reproductive Medicine
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• 제49권2호
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• pp.117-126
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• 2022

Objective: This study examined whether the addition of triple antioxidants (3A)-10 µM acetyl-L-carnitine, 10 µM N-acetyl-L-cysteine, and 5 µM α-lipoic acid-in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa. Methods: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated. Results: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively). Conclusion: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa.

• Clinical and Experimental Reproductive Medicine
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• 제50권4호
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• pp.262-269
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• 2023

Objective: The aim of this study was to compare semen parameters and sperm DNA fragmentation (SDF) and explore the relationship between semen parameters and SDF between 2 and 7 days of abstinence and a short abstinence period (within 4 hours) in oligozoospermic infertile patients. Methods: Two semen samples were collected from infertile oligozoospermic men (n=34) after an abstinence period of 2 to 7 days and within 4 hours, respectively. Sperm parameters were compared between the two abstinence duration groups, including semen volume, sperm concentration, total sperm count, sperm motility, total motile sperm count (TMSC), morphology, and SDF. Results: The semen volume, concentration, and total sperm count were significantly decreased after 4 hours of abstinence than after 2 to 7 days of abstinence, with median differences of 1.2 mL (p<0.001), 2×10⁶/mL (p=0.011), and 9.6×10⁶/ejaculation (p<0.001), respectively. TMSC was significantly lower after a short abstinence, with a median difference of 4.24×10⁶/ejaculate (p<0.001). However, there were no significance differences in the percentage of motility, the SDF, and the percentage of sperm with normal morphology. Interestingly, volume, concentration, total sperm count, sperm motility, and SDF, but not TMSC, exhibited significant linear correlations between the two abstinence groups in univariate regression analysis, except for TMSC. Conclusion: In oligozoospermic men, the volume, concentration, and total sperm count were significantly lower after a short abstinence period, but without adverse effects on sperm motility and SDF.

• 한국패류학회지
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• 제27권3호
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• pp.273-282
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• 2011

The ultrastructural characteristics of germ cells during spermatogenesis and mature sperm morphology in male Pinctada martensii were investigated by transmission electron microscope observation. The morphologies of the sperm nucleus and the acrosome of this species are the oval shape and cone shape, respectively. Spermatozoa are approximately $47-50{\mu}m$ in length including a sperm nucleus (about $1.24{\mu}m$ in length), an acrosome (about $0.60{\mu}m$ in length), and tail flagellum (about $45-47{\mu}m$). The axoneme of the sperm tail shows a 9+2 structure. In P. martensii in Pteriidae, a special substructure showing a thick and wide triangular shape which is composed of electron-dense opaque material (occupied about 50% of all, the upper part of the acrosomal vesicle), appeared in the upper region (part) of the acrosomal vesicle, while the lower region (part) of the acrosomal vesicle is composed of electron-lucent material. Thus, this special structure, which exist in the upper part of the acrosomal vesicle in P. martensii, is somewhat different from those of other subacrosomal vesicle in other families in subacrosomal vesicles. Therefore, we assume that the existence of a special substructure showing a thick and wide triangular shape in the acrosomal vesicle of the spermatozoon can be used as a key characteristic for identification of P. martensii or other species in Pteriidae in subclass Pteriomorphia. The number of mitochondria in the midpiece of the sperm of this species are five (exceptionally sometimes four), as one of common characteristics appear the same number of mitochondria in the same families of superfamilyies. This species in Pteriidae does not contain the axial rod and satellite fibres which appear in the species in Ostreidae in subclass Pteriomorphia. These characteristics can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or tools.

• 한국패류학회지
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• 제27권2호
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• pp.121-129
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• 2011

The ultrastructural characteristics of germ cell development during spermatogenesis and mature sperm morphology of in male Scapharca subcrenata were investigated by transmission electron microscope observation. Spermatogonia are located nearest the outer wall of the acinus, while spermatocytes and spermatids are positioned near the accessory cells. The accessory cells, which is in close contact with developing germ cells, contained a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in supplying of the nutrients for germ cell development. The morphologies of the sperm nucleus and the acrosome of this species are the oval shape and cone shape, respectively. Spermatozoa are approximately 45-$50{\mu}m$ in length including a sperm nucleus (about $1.30{\mu}m$ in length), an acrosome (about $0.59{\mu}m$ in length), and tail flagellum (about 43-$47{\mu}m$). The axoneme of the sperm tail shows a 9 + 2 structure. As some characteristics of the acrosomal vesicle structures, the right and left basal rings show electron opaque part (region), and also the anterior apex part of the acrosomal vesicle shows electron opaque part (region). These characteristics of the acrosomal vesicle were found in Acinidae and other several families in subclass Pteriomorphia. These common characteristics of the acrosomal vesicle in subclass Pteriomorphia can be used for phylogenetic and taxonomic analysis as a taxonomic key or a significant tool. The number of mitochondria in the midpiece of the sperm of this species are five, as one of common characteristics appear in most species in Arcidae and other families in subclass Pteriomorphia. The acrosomal vesicles of Arcidae species do not contain the axial rod and several transverse bands in acrosome, unlkely as seen in Ostreidae species in subclass Pteriomorphia, These characteristics can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or tools.

• Asian-Australasian Journal of Animal Sciences
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• 제28권12호
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• pp.1713-1720
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• 2015

The present work describes experiments undertaken to evaluate the usefulness of selected physicochemical indices of semen, cell membrane integrity and sperm chromatin structure for the assessment of boar semen sensitivity to processes connected with pre-insemination procedures. The experiments were carried out on 30 boars: including 15 regarded as providers of sensitive semen and 15 regarded as providers of semen that is little sensitive to laboratory processing. The selection of boars for both groups was based on sperm morphology analyses, assuming secondary morphological change incidence in spermatozoa as the criterion. Two ejaculates were manually collected from each boar at an interval of 3 to 4 months. The following analyses were carried out for each ejaculate: sperm motility assessment, sperm pH measurement, sperm morphology assessment, sperm chromatin structure evaluation and cell membrane integrity assessment. The analyses were performed three times. Semen storage did not cause an increase in the incidence of secondary morphological changes in the group of boars considered to provide sperm of low sensitivity. On the other hand, with continued storage there was a marked increase in the incidence of spermatozoa with secondary morphological changes in the group of boars regarded as producing more sensitive semen. Ejaculates of group I boars evaluated directly after collection had an approximately 6% smaller share of spermatozoa with undamaged cell membranes than the ejaculates of boars in group II ($p{\leq}0.05$). In the process of time the percentage of spermatozoa with undamaged cell membranes decreased. The sperm of group I boars was characterised with a lower sperm motility than the semen of group II boars. After 1 hour of storing diluted semen, the sperm motility of boars producing highly sensitive semen was already 4% lower ($p{\leq}0.05$), and after 24 hours of storage it was 6.33% lower than that of the boars that produced semen with a low sensitivity. Factors that confirm the accuracy of insemination male selection can include a low rate of sperm motility decrease during the storage of diluted semen, low and contained incidence of secondary morphological changes in spermatozoa during semen storage and a high frequency of spermatozoa with undamaged cell membranes.

• 한국패류학회지
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• 제27권2호
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• pp.149-157
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• 2011

Some characteristics of the formations of acrosomal vesicles during the late stage of spermatids during spermiogenesis and taxonomical charateristics of sperm morphology in male two species (Saxidomus purpurata and Meretrix petechialis) in the family Veneridae were investigated by electron microscope observations. In two species, the morphologies of the spermatozoa have the primitive type and are similar to those of other bivalves in that it contains a short midpiece with five mitochondria surrounding the centrioles. The morphologies of the sperm nuclear types of S. purpurata and M. petechialis in Veneridae have the curved cylindrical and cylinderical type, respectively. And the acrosome shapes of two species are the same cap-shape type. In particular, the axial filament is not found in the lumen of the acrosome of two species, however, subacrosomal material are observed in the subacrosomal spaces between the anterior nuclear fossa and the acrosomal vesicle of two species. The spermatozoon of S. purpurata is approximately 46-$52{\mu}m$ in length, including a curved sperm nucleus (about $3.75{\mu}m$ in length), a long acrosome (about $0.40{\mu}m$ in length),and a tail flagellum (about 45-$47{\mu}m$ long). And the spermatozoon of M. petechialis is approximately 47-$50{\mu}m$ in length including a slightly curved sperm nucleus (about $1.50{\mu}m$ in length), an acrosome (about $0.56{\mu}m$ in length) and tail flagellum (44-$48{\mu}m$ in length). In two species, the axoneme of the sperm tail flagellum of each species consists of nine pairs of microtubules at the periphery and a pair of cental doublets at the center. Therefore, the axoneme of the sperm tail flagellum shows a 9 + 2 structure. In particular, taxonomically important some charateristics of sperm morphologies of two species in the family Veneridae are acrosomal morphology of the sperm, The axial filament is not found in the acrosome as seen in a few species of the family Veneridae in the subclass Heterodonta. The acrosomal vesicle is composed of right, left basal rings and the apex part of the acrosomal vesicle. In particular, right and left basal rings show electron opaque part (region), while the apex part of the acrosomal vesicle shows electron lucent part (region). These charateristics belong to the subclass Heterodonta, unlikely a characteristic of the subclass Pteriomorphia showing all part of the acrosome being composed of electron opaque part (region). Therefore, it is easy to distinguish the families or the subclasses by the acrosomal structures. The number of mitochondria in the midpiece of the sperm of S. purpurata and M. petechialis in Veneridae are five. However, the number of mitochondria in the midpiece of the sperm in most species of Veneridae in the subclass Heterodonta are four. Therefore, the number of mitochondria of the sperm midpiece of two species are exceptionally 5, and it is only exceptional case in the species in Veneridae in the subclass Heterodonta. Except these cases, the number of mitochondria in the sperm midpiece in all families in the subclass Heterodontaare are 4, and now widely used in taxonomic analyses.

• Clinical and Experimental Reproductive Medicine
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• 제46권2호
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• pp.67-75
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• 2019

Objective: Sperm cryopreservation has been widely used in assisted reproductive technology, as it offers great potential for the treatment of some types of male infertility. However, cryopreservation may result in changes in membrane lipid composition and acrosome status, as well as reductions in sperm motility and viability. This study aimed to evaluate sperm DNA fragmentation damage caused by conventional freezing using the sperm chromatin dispersion test. Methods: In total, 120 fresh human semen samples were frozen by conventional methods, using SpermFreeze Solution as a cryoprotectant. Routine semen analysis and a Halosperm test (using the Halosperm kit) were performed on each sample before freezing and after thawing. Semen parameters and sperm DNA fragmentation were compared between these groups. Results: There was a significant decrease in sperm progressive motility, viability, and normal morphology after conventional freezing (32.78%, 79.58%, and 3.87% vs. 16%, 55.99%, and 2.55%, respectively). The sperm head, midpiece, and tail defect rate increased slightly after freezing. Furthermore, the DNA fragmentation index (DFI) was significantly higher after thawing than before freezing (19.21% prior to freezing vs. 22.23% after thawing). Significant increases in the DFI after cryopreservation were observed in samples with both normal and abnormal motility and morphology, as well as in those with normal viability. Conclusion: Conventional freezing seems to damage some sperm parameters, in particular causing a reduction in sperm DNA integrity.

• 대한의용생체공학회:의공학회지
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• 제17권1호
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• pp.25-32
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• 1996

Hough변환을 이용하여 정자의 행태학적 특성을 분석하는 새로운 방법을 개발하였다. 이 방법은 정자의 머리부분이 주위의 배경과 밝기의 차이를 갖는다는 특성과 함께 그 형태가 타원에 근접한다는 특성에 근거하고 있다. 정자의 머리부분은 타원을 나타내는 5개의 파라메터로 표현되고, 최적의 파라메터 값이 반복적인 Hough 변환에 의하여 검출된다. 일차적으로 정자가 있는 영역을 선택한다. Hough변환을 이용한 계산을 가능하게 하기 위하여 Hough변환된 공간의 크기를 최소한으로 줄이고, 검출된 오차가 최소화되는 방향으로 검사영역을 이동 시켜가며 최적의 파라메터들을 도출한다. 형태학적 특성을 나타내는 지표들은 정자의 형태를 나타내는 타원의 다섯개 파라메터들로부터 계산된다.