• Development and Reproduction
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• v.13 no.4
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• pp.321-327
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• 2009

This study aimed to examine the testis toxicities of metal compound, manganese (Mn), which may be generated as mist or fume in the industrial sites. As well as serum prolactin (PRL) concentration was analyzed because Mn accumulation in basal ganglia up-regulates serum PRL and hyperprolactinemia consecutively induces the testis toxicity. Male F344 rats were divided into the 4 groups (2 controls and 2 Mn treated groups, n=10) on the basis of the test condition (inhalation, Mn $1.5mg/m^3$ or not) and treatment period (for 4-weeks and 13-weeks). The treatment time was 6 hr. a day, 5 days a week for the whole body. Basic tests including changes in body weight, feed rate were observed. Blood and testis Mn concentration, and testis toxicity test such as the number and deformity test of sperm were also observed. Serum PRL level was analyzed by ELISA to certify the relationship between the Mn induced increase of the serum PRL level and sperm production. Blood and testis Mn concentrations were significantly and dose-dependently increased. Sperm count was decreased in Mn-treatment groups than control in a treatment time dependent manner. Morphological analysis of cauda epidydimal sperm showed that the frequencies of morphologically abnormal sperms such as bent tail and small head were increased in the both Mn-treatment groups than control. A significant increase in serum PRL levels was found in response to Mn treatment but it was not hyperprolactinemia range. These results suggest that treatment of Mn up-regulates the serum PRL concentration and induces the testis toxicity. The No Aversed Effect Level (NOAEL) of inhaled Mn on the male rat testis may be under the $1.5mg/m^3$.

• Development and Reproduction
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• v.5 no.1
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• pp.53-57
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• 2001

Sperm capacitation and acrosome reaction (AR) have been known to be Ca$^{2+}$-dependent events. Sperm capacitation accompanies with cholesterol efflux fiom plasma membrane, that eventually stimulates AR. However, whether the AR mediated by cholesterol efflux is Ca$^{2+}$ dependent has not been verified yet. Recently, methyl beta cyclodextrin (MBCD) was found to evoke AR by stimulating the cholesterol efflux fiom sperm membrane. In the present study, we examined the requirement of Ca$^{2+}$ in the MBCD-induced AR. During incubation of sperm in the bicarbonate buffered media MBCD increased AR in a dose-dependent manner regardless of the Ca$^{2+}$ presence. In the presence of low molar concentration of Ca$^{2+}$ (100 ${\mu}$M), MBCD-induced AR was slightly increased compared to Ca$^{2+}$-free condition. In the absence of Ca$^{2+}$ supplement, spontaneous AR was slightly increased during the incubation but inhibited by 100 ${\mu}$M EGTA. MBCD potentiated AR even the presence of EGTA. However, EGTA attenuated MBCD-induced AR, suagesting the functional involvement of intracellular Ca$^{2+}$ in the MBCD-induced AR. Taken together, it was suggested that cholesterol efflux from the sperm plasma membrane was sufficient for induction of AR even in the absence of extracellular Ca$^{2+}$and that a condition permissive for mobilization of intracellular Ca$^{2+}$ is important for MBCD-induced AR.

• Applied Microscopy
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• v.29 no.2
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• pp.177-187
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• 1999

Ultrastructure of gametes in the three-spine stickleback, Gasterosteus aculeatus aculeatus was observed, utilizing light, scanning and transmission electron microscopes. The egg of three-spine stickleback is spherical and demersal type. The eggs are highly adhesived to each other but not to substrates. There are many oil droplets in vitelline membrane. The outer surface of egg envelope is arranged by mushroom-like structures and pore canals. The egg have a micropyle, sperm entry site, in the area of the animal pole. The egg envelope consists of three layers, an outer layer with high electron density, a middle layer consisting two layers and an inner layer consisting of 16 to 20 layers. In the fertilized egg envelope, the molecular weights of these components ranged from 14 kDa to 205 kDa. The molecular weights of nam protein bands are 19.4 kDa, 36.7 KDa, 39.4 kDa, 42.9 kDa, 46.1 kDa and 53.0 kDa. The head of spermatozoa is spherical shape and the acrosome is absent. The mitochondria in midpiece are arranged from one to three layers and separated from the axoneme by the cytoplasmic canal. The tail has two lateral fins and the axoneme is of the 9+2 structure.

• Applied Microscopy
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• v.29 no.2
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• pp.241-253
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• 1999

Fertilization pattern of north temperate bats is known to be unique for their sperm storage in the female reproductive tract during hibernation (e.g. Korean greater horseshoe bats). They copulate in fall but their ejaculated spermatozoa survive until the next spring. In another words they can persist to survive during long hibernation under the cold condition $(8\sim13^{\circ}C)$ and are to be fertilized with the ovum ovulated in the next spring, so called delayed fertilization. The present study was designed to observe morphological and functional changes of spermatozoa plasma membrane of the bats, hamsters which are hibernators, and mice which are non-hibernators in the room and the cold (bat-hibernation) temperatures and to confirm influence of the temperature on spermatozoa; survival rate, acrosome reaction rate, protein distribution, $Na^+-K^+-ATPase$ activities and scanning electron microscopic histochemistry. Based on the experimental results obtained in the present study, there were no significant morphological and functional differences in the spermatozoa plasma membrane in both the room and cold (bat-hibernation) temperatures and such an absence of difference suggests that the spermatozoa plasma membrane might play a pertinent role as a protector for consistent fertilization during and after hibernation.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.305-315
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• 2001

In this study, the production of transgenic embryo was attempted by microinjection or round spermatid cultured with foreign DNA. At first, the expression of haploid spermatids specific gene, mTP1 in mouse and hPrm2 in hamster spermatids were investigated by RT-PCR method in testes of young mice and hamster testis. The specific gene expression first appeared at 18 days post partum (dpp) in mice spermatid and 20 dpp in hamster spermatid. Therefore, the round spermatids isolated from 17 dpp mice and 19 dpp hamster were used for the introduction of foreign EGFP gene into haploid round spermatids. For the introduction of EGFP gene haploid round spermatids suspended in medium including EGFP gene were treated with a different electric field strength at 0.11, 0.18 and 0.44 ㎸/cm. After electrical stimulation, viability of testicular sperm cells and 67.6%, 66.4% and 49.9%, in mice and 62.6%, 57.9% and 27% in hamster, respectively. These values were significantly lower than those of non-treated control groups 80.5% in mouse and 69.1% in hamster After 72 hrs culture, the highest expression rate of EGFP gene, 28.5% in mice and 32.1% in hamster were obtained from tile spermatogenic cells electroporated by the field strength or 0.18 ㎸/cm. Then, the ability of fertilization and embryonic development of haploid spermatids transfected with foreign EGFP gene were estimated by the microinjection of spermatids into hamster oocytes. The Irate pronuclear formation rate (77.5%) was lower than non-treated control (80%), and the cleavage rate of the treated group (58.8%) was lower than control (65%). To prove the foreign EGFP integration in hamster embryos, 2-cell stage hamster embryos were subjected to the observation under the fluorescence microscope, and the PCR analysis. As a result, about 44% of 2-cell embryos were showed the integration of EFGP gene into their genome. Therefore, These results suggest the possibility to produce transgenic hamsters by microinjection of haploid spermatid transfected with foreign DNA.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.43-49
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• 2001

This study was performed to elucidate the effects of copper poisoning on the reproductive organ of rats. After consecutive oral administrations of copper sulfate, the weights of testis, the numbers and motilities of sperms, organ weights and histological changes of testes were compared between control and experimental groups. 1. Testis weights of 1,000, 2,000 or 4,000 ppm/kg of copper sulfate-administrated rats gradually decreased compared with control group, and the values no significant changes. 2. The sperm numbers of 1,000, 2,000 or 4,000 ppm/kg of copper sulfate-administrated rats were lowered in dose dependent manners than those of control group's and the values no significant changes. 3, The motilities sperms of 1,000, 2,000 or 4:000 ppm/kg of copper sulfate-administrated rats decreased significantly in dose-dependent manners compared with those of control group's. 4. The weights of livers and kidneys of 1,000, 2,000 or 4,000 ppm/kg of copper sulfate-administration rats decreased or increased. 5. Necrosis of hepatocytes around the central veins and infiltrations of fine granules-harboring macrophages in periportal and interstitial tissues were found out in the livers of copper sulfate-administrated rats. The Bowman's capsule and tubules of kidneys were filled with hyaline material.

• Biomedical Science Letters
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• v.4 no.1
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• pp.27-33
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• 1998

This study has been designed in order to examine a physiological role of $Ca^{2+}$ which has been known as an essential factor for capacitation, to confirm whether the enzyme activity of $Ca^{2+}$-ATPase on capacitation is important or not, and to clarify relationship between various levels of the $Ca^{2+}$ concentration and $Ca^{2+}$-ATPase which has been known to be an important factor of the plasma membranes. In the present study applying quercetin, a $Ca^{2+}$-ATPase inhibitor, the enzymatic effect of $Ca^{2+}$-ATPase on capacitation was found to be remarkable: a significant increase of the transition from the original type (type A) to the type B and the type AR of the spermatozoa. This finding suggests that $Ca^{2+}$-ATPase plays an important role in the efflux and the influx of the $Ca^{2+}$ which has been known to be an essential factor the capacitation and acrosome reaction, and that the inhibitory action of the $Ca^{2+}$-ATPase might be a prerequsite step toward the acrosome reaction. The conclusion reached can be deduced as follows: increment of the intracelluar $Ca^{2+}$ concentration occurred by controlling the slope of $Ca^{2+}$ concentration through $Ca^{2+}$-ATPase activities in both the intra- and extracelluar fluid may be an important procedure for capacitation and acrosome reaction, and ultimately for fertilization of the spermatozoa and the ova.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.127-131
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• 2001

The Jeju horse has been raised for centuries in Jeju island. Recently, as the number of this indigenous horse has been dropped dramatically, this breed became Natural Monument #347 to conserve and multiply this endangered breed. To provide the basic information for AI, sexual activity and semen characteristics in Jeju horse were investigated. Jeju horse semen was collected using Missouri style artificial vagina from fertile stallion.\\`she number of mount per ejaculation was 2..3$\pm$1.8, and the ejaculation time was 27.0$\pm$12.5 seconds. The total volume and gel-free volume of semen was 47.8$\pm$26.7 ml and 42.7$\pm$27.4ml, respectively, and the concentration of sperm and the total number of spermatozoa per ejaculation was 200.7$\pm$112.9$\times$10$^{6}$ ml and 7.6$\pm$3.9$\times$10$^{9}$ ml, respectively. The percentage of motile sperm and the number of live spermatozoa per ejaculation was 75.0$\pm$18.2% and 6.1$\pm$3.4$\times$10$^{9}$ ml, respectively, and the pH of gel-free semen was 7.3$\pm$0.2. The total percentage of abnormal sperm was 31.5%, and the percentage of sperm with abnormal head, midpiece and tail was 9.5$\pm$11.7%, 7.0$\pm$4.0% and 15.0$\pm$15.0%, respectively.

• Korean Journal of Agricultural Science
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• v.28 no.2
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• pp.85-91
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• 2001

This study was carried out to investigate the effects of liquid boar semen on reproductive performance in swine artificial insemination. Many factors, which were breeds, time of insemination, breeding season, sperm per dose etc, have been tried to improve reproductive efficiency. Boars were raised at Swine Artificial Insemination Center in National Livestock Research Institute, Sunghwan, Chungnam, Korea. This experiment was carried out from 1995 to 2000. There were no differences in the fertility results compared with 3 breeds (Landrace, Yorkshire and Duroc), frequencies of artificial insemination (double and triple) per estrus cycle and different seasons by using liquid boar semen. There were no significant differences in conception rate, farrowing rate and litter size using 4 trials of 3.0, 2.5, 2.0 and $1.5{\times}10^9/80ml$ in liquid boar semen with 70% of motile sperm cells. We confirmed that the sperm number per dose of $1.5{\times}10^9/80ml$ could be used for commercial artificial insemination.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.10a
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• pp.246-247
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• 2000

수서동물 생식소의 해부학적 구조 및 생식세포의 미세구조는 종 특이성, 생식생태 및 계통에 따라 다르다. 일반적으로 이매패류의 정자는 각 분류군에 따라 핵과 첨체의 모양, 첨체의 axial rod, 미토콘드리아 수, 편모의 수 및 형태 등에서 다양한 특징을 보인다 (Morse and Zardus, 1997). Anadara trapez 등이 속하는 돌조개과 (Arcidae) 패류들의 정자 두부는 난형이며, 첨채에 axial rod가 없고, 중편에 4개의 미토콘드리아를 가진다. (중략)