This experiment was conducted to investigate the effect of the cysteine and glutathione on the motility index and morphology of human spermatozoa at the sperm processing in vitro. After treating the sperm with medium containing cysteine and glutathione, we measured the motility index and morphology at 0.5 h and 24 h. 1. Following the sperm culture for 0.5 h after treating the sperm with the medium containing 0, 1, 5, 10 mM cysteine, curvilinear velocity (VCL) was significantly (p<0.05) higher in control than that in all treatments. And straight-line velocity (VSL) was high at 1 mM and average path velocity (VAP) was low at 5 mM and 10 mM. But the motility (MOT) and morphology (NOM) were not different between control and all treatments. Following the sperm culture for 24 h, the MOT was significantly high in treatment groups (58.9, 74.4 and 62.3%), compared with that in control(28.7%) and the VCL was also high in treatment groups (31.4, 37.9, and 34.0 ${\mu}{\textrm}{m}$/s), compared with that in control (21.3 ${\mu}{\textrm}{m}$/s). The VSL (18.4, 21.7, and 18.9 ${\mu}{\textrm}{m}$/s) was significantly higher than control (10.7 ${\mu}{\textrm}{m}$/s) and the VAP (20.3, 24.7, and 21.4 ${\mu}{\textrm}{m}$/s) in treatments was also compared with that in control (12.6 ${\mu}{\textrm}{m}$/s). The NOM was not difference between control and treatments. 2 Following the sperm culture for 0.5 after treating the sperm with the medium containing 0, 1, 5, 10 mM glutathione, the MOT, VCL, VSL, VAP, and NOM were not different between control and treatments. Following the sperm culture for 24 h, the MOT was higher in treatment groups (82.9, 83.6, 83.4%) than in control (51.1%) and the VCL was higher in treatment groups (50.9, 51.3, and 49.4 ${\mu}{\textrm}{m}$/s) than control (34.1 ${\mu}{\textrm}{m}$/s). The VSL was also higher in treatment (17.1 ${\mu}{\textrm}{m}$/s) and the VAP was also higher in treatment groups (30.1, 32.5, and 29.7 ${\mu}{\textrm}{m}$/s) than in control (19.8 ${\mu}{\textrm}{m}$/s). The NOM was not different between control and treatments.
Swim-up and Wang's tube system are known methods of motile sperm selection and aerobic bacterial removal from the raw semen. This study was designed to evaluate the recovery rate of motile sperm, % normal morphology of sperm, the efficiency of bacterial removal after sperm preparation by the above two methods. The results were as follows. 1. There was more significant increase of sperm concentration in preparation by swim-up than Wang's tube (p<0.05). The concentration of sperm by swim up was changed from $82.5{\times}10^6/ml$ to $6836{\times}10^6/ml$, and Wang's tube was changed from $82.5{\times}10^6/ml$ to $36.0{\times}10^6/ml$. 2. There was significant increase in sperm motility after preparation by two methods in comparison with initial sperm motility (p<0.05), but no statistical difference between two methods was noted. The % motility of sperm by swim-up was increased from 66.1% to 95.7% and Wang's tube from 66.1% to 98.1%. 3. There was significant increase of % normal morphology of sperm in the samples prepared by two methods (p<0.05), from 49.2% to 85.3% in swim-up and from 49.2% to 92.1% in Wang's tube, but there was no statistical significance between them. 4. There was no bacterial growth in aerobic culture after preparation by two methods.
This experiment was carried out to investigate the effect of bovine serum ablumin (BSA), sugars, glycerol equilibration time, straw size and thawing method on the survival index and the morphology of frozen boar spermatozoa. The results obtained were summarized as follow: 1. When the semen frozen in BF5 dilutor as pellet form was thawed in BTS at 37$^{\circ}$and 50$^{\circ}C$, BF5 dilutor with fructose showed higher sperm survival index than that with dextrose, however, when the semen was thawed on dry test tube at 37$^{\circ}C$, BF5 dilutor with sucrose showed higher sperm survival index than with other sugars. 2 When the semen forzen in BF5 dilutor with straw and thawed at 37$^{\circ}C$, BF5 dilutor with dextrose showed higher sperm survival index than those with other sugars, and there was no difference in sperm survival index between 0.5 and 1.0 ml straws. 3. The sperm survival index of frozen sperm was significantly (P<0.05) improved due to addition of BSA (0.05%) to BF5 dilutor. 4. When the extended semen with BF5 dilutor contatining 0.01 to 0.05% of BSA was frozen in the straw, the semen without glycerol equilibration showed significantly (P<0.05) higher sperm survival index than those with 2, 4 and 6 hrs glycerol equilibration time. 5. The sperm frozen in BF5 dilutor with dextrose or fructose, sucrose and raffinose showed 77 to 88% in normal acrosome rate and no difference among sugars. 6. The frozen semen showed lower normal acrosome rate than the first and second diluted semen, whereas the frozen semen showed higher swollen, damaged and missing acrosome rate than the first and second diluted semen. 7. Damaged and missing acrosome rate of sperm head due to freezing was somewhat inhibited by addition of BSA (0.01 to 0.05) to the BF5 dilutor.
The ultrastructural characteristics of germ cell differentiations during spermatogenesis and mature sperm morphology in male $Atrina$ ($Servatrina$) $pectinata$ were evaluated via transmission electron microscopic observation. The accessory cells, which contained a large quantity of glycogen particles and lipid droplets in the cytoplasm, are assumed to be involved in nutrient supply for germ cell development. Morphologically, the sperm nucleus and acrosome of this species are ovoid and conical in shape, respectively. The acrosomal vesicle, which is formed by two kinds of electron-dense or lucent materials, appears from the base to the tip: a thick and slender elliptical line, which is composed of electron-dense opaque material, appears along the outer part (region) of the acrosomal vesicle from the base to the tip, whereas the inner part (region) of the acrosomal vesicle is composed of electron-lucent material in the acrosomal vesicle. Two special characteristics, which are found in the acrosomal vesicle of A. ($S$) $pectinata$ in Pinnidae (subclass Pteriomorphia), can be employed for phylogenetic and taxonomic analyses as a taxonomic key or a significant tool. The spermatozoa were approximately $45-50{\mu}m$ in length, including a sperm nucleus (about $1.43{\mu}m$ in length), an acrosome (about $0.51{\mu}m$ in length), and a tail flagellum (about $46-47{\mu}m$). The axoneme of the sperm tail evidences a 9+2 structure.
Objective: This study was conducted to examine influence of skimmed milk-based extender (SM), INRA 96 extender and BotuSemen Gold extender on parameters of stallions' ejaculate during storage. Methods: In this study, 14 stallions between 4 and 20 years of age were monitored. Total and progressive motility, viability and morphology of sperm were evaluated at time intervals of 24, 48, and 72 hours after collection. Results: The total motility, progressive motility, and values of sperm with normal morphology were significantly higher in the INRA 96 and BotuSemen Gold extenders than in the SM (p<0.01). The sperm viability differed significantly in all extenders (p<0.01). The highest value of sperm viability was in INRA 96 (64.69%±0.67%) and lowest in SM (59.70%±0.81%). The highest differences occurred at 72 hours of storage. Values of total motility, progressive motility and sperm viability decreased over time (p<0.01). In case of sperm morphology there was no statistically significant decrease between 48- and 72-hour time intervals. Conclusion: It can be concluded that the extenders with a chemically defined composition have shown better indicators of insemination capabilities in ejaculates than the SM. The BotuSemen Gold extender is a suitable alternative to the INRA 96, when used within 48 hours; after 72 hours of storage, however, the INRA 96 showed a higher share of viable spermatozoa.
Hakki Uzun;Merve Huner;Mehmet Kivrak;Ertan Zengin;Yusuf Onder Ozsagir;Berat Sonmez;Gorkem Akca
Clinical and Experimental Reproductive Medicine
/
제51권1호
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pp.48-56
/
2024
Objective: This study investigated the relationship of anthropometric and metabolic risk factors with seminal and sex steroidal hormone parameters in a screened population of healthy males. Methods: The participants were healthy young men without chronic or congenital diseases. The body composition parameters that we investigated were measured weight, height, and waist circumference (WC), as well as bioelectrical impedance analysis. Semen samples were analyzed for semen volume, sperm concentration, sperm motility and morphology, seminal pH, and liquefaction time. Biochemistry analysis, including glucose and lipid metabolism parameters, was conducted on fasting blood samples. Testicular volume was calculated separately for each testis using ultrasonography. Results: Body mass index exhibited an inverse association with total sperm count. WC showed negative correlations with numerous seminal parameters, including sperm concentration, total sperm count, sperm morphology, and follicle-stimulating hormone levels. The basal metabolic rate was associated with seminal pH, liquefaction time, and sperm motility. WC, fat mass percentage, and triglyceride levels exhibited negative correlations with sex hormone binding globulin. The measures of glucose metabolism were associated with a greater number of seminal parameters than the measures of cholesterol metabolism. C-reactive protein levels were inversely associated with sperm concentration and total sperm count. Conclusion: Anthropometric and metabolic risk factors were found to predict semen quality and alterations in sex steroidal hormone levels.
Septin4 belong to a family of polymerizing GTP-binding proteins that are required for many cellular functions, such as membrane compartmentalization, vesicular trafficking, mitosis, and cytoskeletal remodeling. Since, Septin4 is expressed specifically in the testis, we aimed to determine the association between Septin4 gene expression with sperm quality, DNA damage, and stress oxidative level in infertile patients. The present study included 60 semen samples that grouped into three groups: normozoospermia (n=20), asthenozoospermia (n=20), astheno-teratozoospermia (n=20). Initially, semen parameters were analyzed by using the World Health Organization protocol. The mRNA expression of Septin4 in sperm was examined using reverse transcription-polymerase chain reaction. Oxidative stress markers, i.e., total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde, were determined by ELISA kit. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm mitochondrial membrane potential (MMP). However, it showed significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels. In conclusion, Septin4 gene expression provides clinical useful information for the diagnosis of male infertility. It might be a marker for discrimination between fertile and infertile patients. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm MMP. However, it shows significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels.
The present study was undertaken to clarify the role of TEST-Yolk Buffer(TYB) as a factor for the improvement of human sperm fertility potential. We examined the effects of low temperature capacitation using TYB on sperm motility (%), motility pattern, normal morphology, true acrosome reaction, sperm penetration assay and human in vitro fertilization. Comparing the TYB method and swim-up method, the sperm motility(%) of selected sperm was not significantly different, but statistically significant differences were found in curvilinear velocity, linearity, lateral head displacement, normal morphology(%) and true acrosome reaction(%)(p<0.05). Results obtained from the sperm penetration assay demonstrated that the penetration index and penetration rate were increased significantly(p<0.05) when the spermatozoa were incubated in TYB, as compared with swim-up method. And fertilization of intact human oocytes was more succesful when spermatozoa were pretreated with TYB at $4^{\circ}C$ for 48 hours as compared with swim-up method. Our results show that TYB method have advantages in terms of enhancement of sperm hyperactivation, increased true acrosome reaction, increased ability to penetrate zona-free hamster ova and augmented fertilization of human oocytes, suggesting that TYB is superior in its ability to preserve sperm motility and fertilizing ability.
Kim, Jin-Hee;Chung, Ee-Yung;Lee, Ki-Young;Choi, Moon-Sul;Seo, Won-Jae;Kim, Sung-Han
한국패류학회지
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제26권4호
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pp.311-316
/
2010
Spermatid differentiations during spermiogenesis and mature sperm ultrastructure in male Crassostrea nipponica were investigated by transmission electron microscope observations. The morphology of the spermatozoon of this species has a primitive type and is similar to those of other bivalves. Mature spermatozoa consist of broad, cap-shaped acrosomal vesicle and an axial rod in subacrosomal materials on an oval nucleus showing deeply invaginated anteriorly, two triplet substructure centrioles surrounded by four spherical mitochondria, and satelite fibres, which appear near the distal centriole. The acrosomal vesicle of spermatozoa of C. nipponica resemble to those of other investigated ostreids. Especially, two transverse bands (stripes) appear at the anterior region of the acrosomal vesicle, unlikely 2-3 transverse bands (stripes) in C. gigas. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm morphology in the resolution of taxonomic relationships within the Ostreidea. The sperm is approximately $48-50{\mu}m$ in length including an oval sperm nucleus (about $1.0{\mu}m$ in length and $1.41{\mu}m$ in width), an acrosome (about $0.48{\mu}m$ in length and 0.30 in width) and tail flagellum ($46-48{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9 + 2 structure. These morphological charateristics of acrosomal vesicle belong to the family Ostreidae in the subclass Pteriomorphia.
This study was carried out to obtain informations regarding the effect of N-acetyl-D-glucosamine in the LEY (lactoseegg yolk) diluent according to incubation time in 5 ml maxi-straw and the effects of freezing rate, thawing temperature and thawing time in the LEN (lactose-egg yolk and N-acetyl-D-glucosamine) diluent on acrosome morphology and motility of frozen-thawed boar sperm. The study showed that the LEN diluent was higher post-thaw NAR (normal apical ridge) acrosome than the LEY diluent for 0.5 h incubation at 37$^{\circ}C$. However, there were no differences between the LEN and LEY diluents on post-thaw sperm motility according to incubation time. The straws frozen from 5.0 cm (20$^{\circ}C$/min) to 17.0 cm (1$^{\circ}C$/min) above the liquid nitrogen surface did not show any significant differences on post-thaw sperm motility. However, the straws frozen above 5.0 cm from the liquid nitrogen surface were higher NAR acrosome than those frozen above 17.0 cm. The post-thaw percentages of motile sperm and NAR acrosome were significantly higher (p<0.05) for the maxi-straws submerged for 40 or 45 sec in a 52$^{\circ}C$ water bath than for 30, 35, 50 or 55 sec. The mean sample temperatures of maxi-straws after 40 or 45 sec submersion were 20.7 or 26.4$^{\circ}C$. In conclusion, the sample temperature of the thawed semen was very important for post-thaw sperm survival in the LEN diluent of 5 ml maxi-straw. When the temperature of the thawed semen was 20.7$^{\circ}C$, the percentages of motile sperm and NAR acrosome were highest.
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