• Molecules and Cells
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• v.28 no.5
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• pp.441-446
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• 2009

During epididymal transit, mammalian sperm acquire selected proteins secreted by the epididymis. We previously showed that a disintegrin and metalloprotease (ADAM) 7 is expressed specifically in the epididymis and transferred to the sperm surface during epididymal transit. Here, we show that mouse ADAM7 secreted to the epididymal lumen is associated with membranous vesicles known as epididymosomes. Furthermore, we found that ADAM7 can be transferred directly from epididymal vesicles to sperm and that it is an integral plasma membrane protein in sperm. Thus, our study provides new information regarding the unique mode of secretion and interaction of ADAM7 during the epididymis-to-sperm transfer process.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.247-253
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• 1997

The objective of this study was to examine the fertilization and embryonic development rates of porcine oocytes matured in vitro according to the morphological normality of epididymal sperm in porcine IVF. The results obtained in this experiment were summarized as follows : 1. When the ratio of morphological normality of epididymal sperm was divided into the three groups with $\leq$ 10%, 10-30% and $\geq$ 50%, the fertilization and embryonic devel-opment rates of 능50% group (64%. 26%) w were significantly higher than those of other two groups ($\leq$10%: 27%, 6% and 10-30%: 36%. 5%) (p$\leq$50% group of morphologically normal epididymal sperm was adjusted to 100% (5 x 10$^5$ cells/ml), the fertilization a and development rates (63%, 27%) of epidymal sperm were similar to those of frozen-thawed ejaculated sperm (56%, 35%). 3. Also, when the fertilization and development rates of epididymal sperm were analyzed according to the oocyte : sperm ratio (1:6000, 1: 6650, 1: 7700, 1: 10000), the fertilization and development rates indicated high, in 1:6000 (68%, 32%) and 1:6650 (89%, 31%), the ratio of oocyte: sperm. Therefore, when the percentage of morphological normality of epididymal sperm is more than 50, the embryonic development a can be obtained similar to that of frozen thawed ejaculated sperm. Also, these result suggested that the morphological evaluation of normal sperm in porcine IVF using epididymal sperm sho비d be prerequisite for the more effective embryonic developm ment.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.275-279
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• 2009

The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

• Korean Journal of Poultry Science
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• v.27 no.1
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• pp.63-71
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• 2000

The testis is an extremely heterogeneous organ, containing numerous compartments types. Morphometric studies were performed of 3 avian species (pigeon, pheasant and chicken) to determine volume density absolute volume, numerical density, total number of serminiferous tubule components, and sperm production, especially those related to the Sertoli cell, and to make comparisons among the species. Volume density of seminiferous tubule components per testis was determined by point counting method. Testis volume and sperm production were measured by routine techniques. Numerical density (the number of cells per unit volume of testis) of seminiferous tubule components per testis was determined by morphometry (Floderus method). The volume density of seminiferous tubules per testis was 91.58, 92.18 and 94.21% in pigeon, pheasant, and chicken, respectively. The volume density of spermatogonium, spermatocyte, spermatid, spermatozoon, and Sertoli cell did not produce significant changes in the three species. The absolute volume of spermatogonium, spermatocyte, spermatid, and Sertoli cell showed significant changes in the three species (p<0.05). The average volume of Sertoli cell ranged from 758.34(pheasant) to 1,212.9 ㎛$^3$(chicken) and was not significantoy different in the three species(p>0.05). The number of Sertoli cells per testis showed significant differences in the three species : 34.52 $\times$10(sup)6, 186.82$\times$10(sup)6, 810.62$\times$10(sup)6 in pigeon, pheasant, and chicken, respectively(p<0.05). The sperm production was significantly different in the three species : 3,018$\times$10(sup)6, 993.9$\times$10(sup)6, and 8.9$\times$10(sup)6 in chicken, pheasant, and pigeon, respectively(p<0.05). These results suggest that number of Sertoli cells may be more important than Sertoli cell size in explaining the difference in sperm production among the three species.

• Biomedical Science Letters
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• v.23 no.4
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• pp.406-410
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• 2017

Curcumin is known as a natural antioxidant, decreasing oxidative stress in animal cells. Generally, oxidative stress induces reactive oxygen species in sperm and leads to decreased sperm characteristics in pigs. Therefore, this study investigated the influence of curcumin on sperm motility, viability, mitochondrial activity and plasma membrane integrity in pigs. Curcumin (0, 5 and $10{\mu}M$) was treated in boar semen, which were incubated for 9 hours in $37^{\circ}C$. Then, motility, viability, mitochondrial activity, plasma membrane integrity of sperm was analyzed every 3 hours. In the results, sperm motility was significantly increased by $5{\mu}M$ curcumin after 3 and 9 hours after incubation, and viability was significantly higher in $5{\mu}M$ curcumin treatment at 3 hours (P<0.05). Similarly, sperm mitochondrial activity and plasma membrane integrity were significantly increased by $5{\mu}M$ curcumin at 3, 6 and 9 hours after incubation (P<0.05). There results suggest that curcumin improve sperm characteristics such as motility, viability, mitochondrial activity, and plasma membrane integrity, and may exert a positive effect on sperm fertility in pigs.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.229-234
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• 2002

This study was carried out to investigate the general characteristics such as concentration sperm motility and abnormality of sperm on the whole epididymal semen(EWS), RSP-S(removed seminal plasma by saline) and RSP-T(removed seminal plasma by tris-buffer) semen and survival rates after freezing on motility of whole and RSP-S and RSP-T semen and extender containing 2~8% glycerol, and ability of frozen-thawed sperm to penetrate homologous oocytes. 1. The concentration, motility and abnormality of epididymal WES, RSP-S and RSP-T sperm were 4.25 $\pm$ 0.25($\times$10$^{6}$ Cells/$m\ell$), 3.85$\pm$0.20($\times$10$^{6}$ Cells/$m\ell$), 4.05 $\pm$ 0.28($\times$10$^{6}$ Cells/$m\ell$), 50.55 $\pm$ 2.75%, 67.25 $\pm$ 2.55%, 78.75 $\pm$ 3.55 and 9.45 $\pm$ 2.25%, 37.75 $\pm$ 2.10%, 24.25 $\pm$ 1.55%, respectively. 2. The survival rates of slow and rapid frozen epididymal RSP-S and RSP-T sperm were 35.00 $\pm$ 2.35%, 45.50 $\pm$ 2.15％ and 16.50 $\pm$ 3.55%, 22.55 $\pm$ 3.95%, respectively. The survival rate of epididymal WES and RSP-T sperm after freezing following dilution with tris-buffer containing 2~8% glycerol were 9.25 $\pm$ 1.55%~17.50 $\pm$ 2.50%. 3. The percentage of capacitated and acrosome-reacted sperm prier to culture for fresh and frozen -thawed epididymal RSP-T semen were 45.25 $\pm$ 5.75%, 7.06 $\pm$ 0.25%, 48.20 $\pm$ 6.80% and 13.00 $\pm$ 2.35%, 3.55 $\pm$ 0.85%, 15.50 $\pm$ 1.90%, respectively. The penetration rate the number of sperm per penetrated for fresh and frozen-thawed epididymal RSP-T sperm were 39.25 $\pm$ 4.72%, 34.24 $\pm$ 3.93% and 1.30 $\pm$ 0.33, 1.10 $\pm$ 0.50., respectively.

• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.245-252
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• 1990

This study was carried out to investigate the factors affecting fertilization in vitro of follicular oocytes with frozen-thawed spermatozoa in Korean Native Cattle. The bovine ovaries were obtained at a slaughter house and the follicular oocytes were recovered by aspirating the follicular fluid from the visible follicles of 3~6mm. The bovine oocytes were matured in vitro for 20~24 hours in TCM-199 containing FCS and hormones. The matured oocytes were fertilized in vitro using Percoll-separated frozen-thawed spermatozoa in BO solution. The effects of dilution and fertilization media, capacitating method, concentration of inseminated sperm and time after insemination of fertilization, were observed. The results obtained are summarized as follows : 1. The fertilization rate of frozen-thawed sperm inseminated in BO solution with caffeine and heparin together(56.4%) was higher than that of sperm inseminated in BO solution with either caffeine(10.5%) or heparin(8.9%) and without both caffeine and heparin(0%)(P<0.05). 2. The fertilization rate(56.3%) of frozen-thawed sperm inseminated in BO solution with both caffeine and heparin without preincubation was higher than that of sperm preincubated(2.9%)(P<0.05). 3. The fertilization with high concentration of frozen-thawed sperm(1.4~1.8$\times$107cells/ml) in BO solution containing caffeine and heparin resulted in higher fertilization rate, 76.7%, than the low concentration of sperm(0.8~1.0$\times$107cells/ml), 32.7%(P<0.01). 4. When the oocytes were inseminated with frozen-thawed sperm in BO solution containing caffeine and heparin without preincubation, fertilization rate increased by time and the rates were 5.9, 46.0 and 59.4% at 8, 16 and 24 hours, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.1
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• pp.19-25
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• 2023

Objective: Sleep deprivation (SD) is a common problem in today's stressful lifestyle and have physiological consequences, including reproductive dysfunction and infertility. As an antioxidant, olive oil may be effective in reducing testicular and spermatological damage by decreasing the production of free radicals. Methods: This study investigated the effects of olive oil on sperm quality and testicular structure using stereological methods to assess rats with SD. Results: When comparing SD group to grid floor+distilled water (GR) group, we found that the sperm count and motility, as well as the percentage of slow progressive sperm was significantly lower in SD group (p<0.05), but the percentage of immotile sperm was higher (p<0.01). However, no improvement was observed in sperm count or motility after concomitant treatment of SD group with olive oil. Stereological examinations revealed no significant change in the total volumes of the seminiferous tubules, interstitial tissue, and germinal epithelium in the study groups. Conversely, the total number of testicular cell types was significantly lower in SD group than in GR group. Although the total number of Sertoli and Leydig cells was significantly higher in the S +olive oil group than in the untreated SD group, no significant difference in the total number of other testicular cell types was observed between the two groups. Conclusion: SD potentially induced structural changes in testis that affected sperm count and motility. However, olive oil only improved the total number of Sertoli and Leydig cells in the animals with SD and did not improve sperm count and motility.

• BMB Reports
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• v.52 no.8
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• pp.482-489
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• 2019

Reproductive biotechnology has developed rapidly and is now able to overcome many birth difficulties due to infertility or the transmission of genetic diseases. Here we introduce the next generation of assisted reproductive technologies (ART), such as mitochondrial replacement technique (MRT) or genetic correction in eggs with micromanipulation. Further, we suggest that the transmission of genetic information from somatic cells to subsequent generations without gametes should be useful for people who suffer from infertility or genetic diseases. Pluripotent stem cells (PSCs) can be converted into germ cells such as sperm or oocytes in the laboratory. Notably, germ cells derived from nuclear transfer embryonic stem cells (NT-ESCs) or induced pluripotent stem cells (iPSCs) inherit the full parental genome. The most important issue in this technique is the generation of a haploid chromosome from diploid somatic cells. We hereby examine current science and limitations underpinning these important developments and provide recommendations for moving forward.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.321-326
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• 2018

In this study, two epididymal spermatozoa recovery methods in relation to sperm number, motility, viability and acrosome reaction were examined. Seven bulls were castrated and 7 testicles with epididymides were transferred to the laboratoy. Epididymis in each bull was randomly used for flushing and mincing methods with semen extender (Optixcell, IMV, France). The recovered spermatozoa with adjusted sperm concentration to $40{\times}10^6cells/mL$ was diluted with optixcell and cryopreserved. In experiment 1, the difference in the total number of spermatozoa using flushing and mincing methods was insignificant (2570.0 and $2505.2{\times}10^6cells/mL$, respectively). For experiment 2, the percentage of motile spermatozoa and motility parameters between flushing and mincing methods were studied through the use of sperm class analyzer after frozen-thawing. The percentage of total motile sperm between flushing and mincing methods was almost the same with $89.5{\pm}12.8$ and $91.4{\pm}7.9%$, respectively. The same is the case with experiment 3 wherein the viability and acrosomal integrity of frozen-thawed epididymal spermatozoa by flushing and mincing was insignificantly different. The results from the study showed that both flushing and mincing methods can be used for epididymal spermatozoa recovery in bull.