• The Korean Journal of Physiology and Pharmacology
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• v.18 no.2
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• pp.143-148
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• 2014

Arecoline is a major alkaloid of areca nuts which are widely chewed by southeast Asian and it manifests various toxic effects in different organs of human and animals. In this work, mature mice were treated by vitamins C plus E, arecoline, or both daily for four weeks. The results showed that arecoline significantly increased the levels of serum alkaline phosphatase (ALP), glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and significantly decreased the levels of reduced glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) in the liver tissues. Additionally, the body weight, testis weight, sperm counts, motility and normal sperms also were significantly decreased. The supplement of vitamins C and E can bring the activities of ALP and GPT to normal levels and partially restore the sperm counts compared to the arecoline-treated group but have no other positive effects. In conclusion, the vitamins C and E partially attenuated the arecoline-induced hepatotoxiciy but basically had on protective effects against the arecoline-induced testicular toxicity.

• Korean Journal of Ichthyology
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• v.26 no.2
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• pp.89-98
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• 2014

Eggs development and early life history of the endangered Korean dwarf loach, Kichulchoia brevifasciata (Teleostei: Cobitidae) was investigated to provide basic information regarding biological characteristics and restoration. Adult fish specimens were sampled using a spoon net at Geurnsan-myeon, Goheung-gun, Jeollanam-do, Korea from June to July 2011. Since, spawning characteristics were analyzed, and females were induced to spawn by injecting Ovaprim (0.5 mL/kg) and their eggs were artificially fertilized with sperms by the dry method in the laboratory. Total length of mature female were 46~76 mm with GSI $9.6{\pm}3.77%$, and total length of mature male was 42~52 mm with GSI $3.5{\pm}1.04%$. Sex ratio (♂/♀) was 0.10, and there were no secondary sexual characteristics. The number of mature eggs was averaged $60{\pm}28.7$ per female. The lemon yellow eggs were slightly adhesive $1.46{\pm}0.07mm$ in diameter. The embryo hatched approximately 66 h after fertilization at $25^{\circ}C$, and the hatched larvae were averaged $5.5{\pm}0.07mm$ in total length (TL). At 6 days after hatching, the larvae averaged $9.0{\pm}0.29mm$(TL) and their yolk sac was completely absorbed. At 17 days after hatching, they entered the juvenile stage and reached $12.6{\pm}0.24mm$ (TL). At 80 days after hatching, the band patterns and external form of the juveniles were similar to those of adults, and they averaged $33.0{\pm}2.19mm$(TL).

• Development and Reproduction
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• v.21 no.3
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• pp.327-333
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• 2017

Gonadotropins are heterodimers consisting an alpha chain ($Cg{\alpha}$) and a beta chain. Interestingly, presence of complicated $LH-{\beta}$ transcripts in rat testis was accidently found; testicular $LH-{\beta}$ transcripts were confined in seminiferous tubules to spermatids, and the translated products were localized in the elongated spermatids. We hypothesized that mouse testis has potential to produce the tissue specific $LH-{\beta}$ with similar structure to the rat testicular forms. To verify our hypothesis, we examined the adult mouse (ICR) testis using RT-PCR and immunohistochemistry. The PCR revealed the presence of the identical products in the reactions for three LH subunit types. The expected product sizes for mouse $Cg{\alpha}$ and $LH-{\beta}$ known as pituitary type were 224 bp and 503 bp, respectively. The testicular type $LH-{\beta}$ products were produced by a primer set based on the rat sequences, with unexpected size of 800 bp. Sequencing revealed that the proximal and distal parts (2-82 and 661- 773 bp, respectively) were homologous to rat testicular $LH-{\beta}$ cDNA, and middle part (83-660 bp) was a unique mouse-specific region. Both $Cg{\alpha}$ and $LH-{\beta}$ positive signals were in the round and elongated spermatids and mature sperms, and the $LH-{\beta}$ signals were more intense. In conclusion, our study demonstrated that the presence and localization of the LH subunits in mouse testis. Further studies will be needed to understand the precise structure and function of mouse testicular LH.

• Reproductive and Developmental Biology
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• v.33 no.1
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• pp.55-61
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• 2009

The epididymis in the male reproductive tract is the site where spermatozoa produced from the testis become mature. The epididymis is divided into 4 different segments, initial segment and caput, corpus, and caudal epididymis, depending upon functional and morphological features. Aquaporins (Aqps) are water channel molecules, which are present in the epididymis and play a major role in removal of epididymal water, resulting in creation of microenvironment for sperm maturation and concentration of sperms. Nandrolone decanoate (ND) is a synthetic anabolic-androgenic steroid, which is used to treat clinical diseases and improve physical ability and appearance. Even though it is well determined that the ND causes the male infertility by affecting the testis, little is known the effect of the ND on the epididymis. The present study was focused to examine the effect of ND at different treatment doses and periods on expression of Aqp1 and Aqp9 genes in the epididymis of pubertal rats. Results showed that mRNA expression of Aqp1 and Aqp9 genes among the parts of the epididymis was differentially regulated by ND treatment doses. In addition, treatment periods of ND caused differential expression of Aqp1 and Aqp9 mRNAs among segments of the epididymis. Therefore, it is believed that male infertility induced by ND could be resulted not only from malfunction of the testis but also from aberrant gene expression of Aqp1 and Aqp9 in the epididymis.

• Applied Microscopy
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• v.39 no.3
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• pp.245-252
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• 2009

The reproductive cells in testes of Microphysogobio yaluensis were investigated using light and electron microscopes. The testis of Microphysogobio yaluensis consisted of numerous testicular cysts contained synchronized cells. Sperms were full in testicular sacs of mature testes. Leydig cells were located among testicular cysts. The nucleus of primary spermatocytes was round and mitochondria were congregated in cytoplasm. The size of secondary spermatocyte was smaller than that of primary spermatocyte and the nucleus of a secondary spermatocyte was round or oval. In spermatids, the nucleus was round and electron-dense. In spermiogenesis, the nucleus was condensed and a flagellum started to be formed. The mitochondria were rearranged along the flagellum. The sperm had a round head, the acrosome was not found and a motile flagellum consisted of an axoneme with a typical 9+2 pattern of microtubule.

• Development and Reproduction
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• v.23 no.2
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• pp.93-99
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• 2019

Nonylphenols (NPs) are widely used industrial materials, and are considered as potent endocrine disrupting chemical. Present study was undertaken to clarify the effect of subchronic low-dose NP exposure to F1 generation male mice. Mice were divided into 2 groups; (1) CON, control animals and (2) NP-50 ($50{\mu}g/L$), animals were treated with NP via drinking water. NP exposures were continuously conducted from parental pre-mating period until the postnatal day (PND) 55 of F1 offsprings. Mice were sacrificed on PND 55 and the tissue weights were measured. The initial body weights (at PND 21) and terminal body weights (PND 55) of the NP-50 animals were significantly lower than those of control animals (p<0.05). NP exposure induced a significant increase in the absolute weight of the testes (p<0.05). Conversely, the NP exposure caused significant decrease in the absolute weights of the epididymis (p<0.01), prostate (p<0.05) and seminal vesicle (p<0.05). Histopathological studies revealed that NP-treated animals exerted decreased seminiferous tubule diameters, reduced luminal area, and lower number of germ cells. Also some sloughing morphologies in the tubules were observed. In the caudal epididymis, fewer mature sperms and swollen epithelial cells were found in the NP-treated group. Our results confirmed that the subchronic low-dose NP exposure altered some male parameters and induced histopathological abnormalities in testis and epididymis of F1 mice. Since the NP dose used in this study is close to the average human daily NP exposure, our results could provide practically meaningful understanding of adverse effect of EDC in human.

• Development and Reproduction
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• v.23 no.3
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• pp.255-262
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• 2019

Previously, we reported negative effects of low-dose nonylphenol (NP) exposure on the reproductive organs of F1 male mice. In the present study was further investigated the endocrine disrupting effect of NP exposure to F2 generation male mice. Mice were divided into 2 groups; (1) CON, control animals and (2) NP-50 ($50{\mu}g/L$), animals were treated with NP via drinking water. NP exposures were continuously conducted from parental pre-mating period until the postnatal day (PND) 55 of F2 offsprings. Mice were sacrificed on PND 55 and the reproductive tissue weights were measured. The initial (at PND 21) and terminal (PND 55) body weights of the NP-50 group animals were not significantly different from those of control group animals. NP exposure fail to induce a significant weight change of the testes, seminal vesicle and prostate except absolute epididymal weight (p<0.05). However, pathohistological studies revealed that NP-treated F2 animals showed evident decrease in seminiferous tubule diameters, reduced luminal area and number of germ cells. Also, sloughing morphologies in the tubules were notable. In the caudal epididymis, fewer mature sperms and swollen epithelial cells were found in the NP-treated group. The present study demonstrated that the subchronic low-dose NP exposure induced pathohistological abnormalities in testis and epididymis of F2 mice, and we assumed that these 'qualitative' changes in reproductive tissues could be derived from the epigenetic modifications such as DNA methylation, histone modification, altered DNA accessibility and chromatin structure. Further studies are needed to achieve a better understanding on the multi- or trans-generational effects of NP on the reproductive health and a human application.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.4
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• pp.253-260
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• 2004

Objectives: Klinefelter syndrome is the most common genetic cause of male infertility and presents with 47, XXY mainly or 46, XX/47, XXY mosaicism. It is characterized by hypogonadism and azoospermia due to testicular failure, however, sporadic cases of natural pregnancies have been reported. With the development of testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI), sperm can be retrieved successfully and ART is applied in these patients for pregnancy. It has been suggested that the risk of chromosome aneuploidy for both sex chromosome and autosome is increased in the sperms from 47, XXY germ cells. Considering the risk for chromosomal aneuploidy in the offspring, preimplantation genetic diagnosis (PGD) could be applied as a safe and more effective treatment option in Klinefelter syndrome. The aim of this study is to assess the outcome of PGD cycles by using FISH for sex chromosome and autosome in patients with Klinefelter syndrome. Materials and Methods: From Jan. 2001 to Dec. 2003, PGD was attempted in 8 cases of Klinefelter syndrome but TESE was failed to retrieve sperm in the 3 cases, therefore PGD was performed in 8 cycles of 5 cases (four 47, XXY and one 46, XY/47, XXY mosaicism). In one case, ejaculated sperm was used and in 4 cases, TESE sperm was used for ICSI. After fertilization, blastomere biopsy was performed in $6{\sim}7$ cell stage embryo and the chromosome aneuploidy was diagnosed by using FISH with CEP probes for chromosome X, Y and 17 or 18. Results: A total of 127 oocytes were retrieved and ICSI was performed in 113 mature oocytes. The fertilization rate was $65.3{\pm}6.0%$ (mean$\pm$SEM) and 76 embryos were obtained. Blastomere biopsy was performed in 61 developing embryos and FISH analysis was successful in 95.1% of the biopsied blastomeres (58/61). The rate of balanced embryos for chromosome X, Y and 17 or 18 was $39.7{\pm}6.9%$. The rate of aneuploidy for sex chromosome (X and Y) was $45.9{\pm}5.3%$ and $43.2{\pm}5.8%$ for chromosome 17 or 18, respectively. Embryo transfer was performed in all 8 cycles and mean number of transferred embryos was $2.5{\pm}0.5$. In 2 cases, clinical pregnancies were obtained and normal 46, XX and 46, XY karyotypes were confirmed by amniocentesis, respectively. Healthy male and female babies were delivered uneventfully at term. Conclusion: The patients with Klinefelter syndrome can benefit from ART with TESE and ICSI. Considering the risk of aneuploidy for both sex chromosome and autosome in the sperms and embryos of Klinefelter syndrome, PGD could be offered as safe and more effective treatment option.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.3
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• pp.199-205
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• 2020

Cryopreservation of semen is useful for animal breeding via artificial insemination (AI). However, the use of frozen-thawed boar semen is limited due to cryodamage. The aim of this study was to investigate the effects of different concentrations of MitoTEMPO (a mitochondria-targeted antioxidant) in lactose-egg yolk (LEY) extenders on kinetic characteristics of frozen-thawed boar sperms. Semen samples were collected from mature Duroc boars (2~3 years old) and cryopreserved in LEY extenders containing 0, 0.5, 5, 50, and 500 μM MitoTEMPO. The kinetic characteristics of frozen-thawed sperms were determined 0 and 30 min after thawing using computer-assisted sperm analysis (CASA). Results indicated that sperm motility immediately after thawing was significantly higher with 5 and 50 μM (50.46±2.71% and 46.96±2.66%, respectively) than with 500 μM MitoTEMPO (35.40±2.95%) (P<0.05). However, there were no significant differences in other kinetic characteristics except motility. In conclusion, the addition of MitoTEMPO to the sperm freezing extender may have a beneficial effect on motility of post-thawed boar semen.

• The Korean Journal of Malacology
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• v.26 no.1
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• pp.51-62
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• 2010

The ultrastructures of germ cells and the accessory cells during spermatogenesis and mature sperm ultrastructure in male Gomphina veneriformis, which was collected on the coastal waters of Yangyang, East Sea of Korea, were investigated by transmission electron microscope observations. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves in that it contains a short midpiece with four mitochondria surrounding the centrioles. Accessory cells are observed to be connected to adjacent germ cells, they contain a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in the supplying of the nutrients for germ cell development, while any phenomena associated with phagocytosis of undischarged, residual sperms by lysosomes in the cytoplasm of the accessory cells after spawning was not observed in this study. The morphologies of the sperm nucleus type and the acrosome shape of this species have a cylindrical and modified long cone shape, respectively. In particular, the axial filaments in the lumen of the acrosome, and subacrosomal granular materials are observed in the subacrosomal space between the anterior nuclear fossa and the beginning part of axial filaments in the acrosome. The spermatozoon is approximately $50-55{\mu}m$ in length including a long sperm nucleus (about $7.80{\mu}m$ in length), an acrosome (about $1.13{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$). 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. Some charateristics of sperm morphology of this species in the family Veneridae are (1) acrosomal morphology, (2) the number of mitochondria in the midpiece of the sperm,. The axial filament appears in the acrosome as one of characteristics seen in several species of the family Veneridae in the subclass heterodonta, unlikely the subclass pteriomorphia containing axial rod instead of the axial filament. As some characteristics of the acrosome structures, the peripheral parts of two basal rings show electron opaque part (region), while the apex part of the acrosome shows electron lucent part (region). These charateristics belong to the family Veneridae in 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 acrosome structures. The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appeared in most species in the family Veneridae.