• Journal of Ginseng Research
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• v.40 no.2
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• pp.185-195
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• 2016

Background: To investigate the effect of pectinase-treated Panax ginseng (GINST) in cellular and male subfertility animal models. Methods: Hydrogen peroxide ($H_2O_2$)-induced mouse spermatocyte GC-2spd cells were used as an in vitro model. Cell viability was measured using MTT assay. For the in vivo study, GINST (200 mg/kg) mixed with a regular pellet diet was administered orally for 4 mo, and the changes in the mRNA and protein expression level of antioxidative and spermatogenic genes in young and aged control rats were compared using real-time reverse transcription polymerase chain reaction and western blotting. Results: GINST treatment ($50{\mu}g/mL$, $100{\mu}g/mL$, and $200{\mu}g/mL$) significantly (p < 0.05) inhibited the $H_2O_2$-induced ($200{\mu}M$) cytotoxicity in GC-2spd cells. Furthermore, GINST ($50{\mu}g/mL$ and $100{\mu}g/mL$) significantly (p < 0.05) ameliorated the $H_2O_2$-induced decrease in the expression level of antioxidant enzymes (peroxiredoxin 3 and 4, glutathione S-transferase m5, and glutathione peroxidase 4), spermatogenesis-related protein such as inhibin-${\alpha}$, and specific sex hormone receptors (androgen receptor, luteinizing hormone receptor, and follicle-stimulating hormone receptor) in GC-2spd cells. Similarly, the altered expression level of the above mentioned genes and of spermatogenesis-related nectin-2 and cAMP response element-binding protein in aged rat testes was ameliorated with GINST (200 mg/kg) treatment. Taken together, GINST attenuated $H_2O_2$-induced oxidative stress in GC-2 cells and modulated the expression of antioxidant-related genes and of spermatogenic-related proteins and sex hormone receptors in aged rats. Conclusion: GINST may be a potential natural agent for the protection against or treatment of oxidative stress-induced male subfertility and aging-induced male subfertility.

• Development and Reproduction
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• v.2 no.1
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• pp.75-80
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• 1998

메기 정자는 그 길이가 약 62.5 \mu m이며 구형의 핵, 짧은 중편 및 꼬리를 ㄱ진 일반적인 메기류 정자의 미세구조적 특징을 나타내었다. 정자는 대부분의 경골어류의 정자에서와 같이 첨체를 가지고 있지 않았으며 염색질은 완전치 농축되어 있었다. 핵와(nuclear fossa)는 약 0.6 \mu m 함입되어 있었고 그 속에 기부 중심립과 말단 중심립의 일부가 포함되어 있었다. 두 중심립은 140 \circ C의 각도로 배열되어 있었으며 말단 중심립에서 9개의 부수체가 언형질막을 향하여 배열되어 있었다. 미토콘드리아는 중편 세포질에서 2층 또는 3층으로 배열되어 있었으며 핵의 후반부와 꼬리의 기부를 둘러싸고 있었다. 꼬리는 축사만으로 구성되어 있었으며 lateral fins는 관찰되지 않았다. 메기 정자의 가장 큰 구조적 특징은 중편 세포질에 구성되어 있는 관구조(tubular structure)이었다. 대부분의 경골어류의 정자는 중편 세포질에 미토콘드리아만을 포함하고 있으나, 메기 정자에서는 중편 세포질의 전반부에 미토콘드리아가 포함되어 있고, 후반부에는 소관이 모여 망상구조를 형성하는 관구조가 잘 발달되어 있었다. 이와 같은 관구조는 현재까지 Characiformes의 정자 이외의 다른 경골어류에서는 보고된 바 없으며 이러한 구조는Characiformes과 메기류의 계통학적 관계를 연구하는데 매우 중요한 형질로 여겨진다. ^u The spermatozoa of Silurus asotus are appoximately 62.5 \mu m in length and relatively simple cells composed of spherical head, a short midpiece and a tail as in most Siluriformes. The ultrastructure of the spermatozoa of S. asotus is characterized by the following features. The nucleus measuring about 1.5 \mu m in length is depressed with a deep nuclear fossa of about 0.6 \mu m in length, two fifth of the nuclear diameter. The fossa contains the proximal centriole and the half of the distal centriole. Two centrioles form an angle of approximately 140 \circ to each other. the nine satellite rays radiate from the outer surface of the distal centriole. the mitochondrea surround the basal nucleus and the axoneme, and are arranged in two or three layers in the postnuclear cytoplasm. The lateral fins are lost in the sperm tail. The most significant feature is manifested in the midpiece. The midpece comprises two parts, the mitochondria and the tubular structure unlike other teleost fishes containing only the mitochondria. The tubular structure was reported only in the spermatozoa of Citharinus belonging to the characiformes of teleost fishes. Thus it is considered to be a good characteristics for the study of phylogenetic link between Siluriformes and Characiformes.

• Development and Reproduction
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• v.2 no.1
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• pp.101-109
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• 1998

Spermatogenesis and reproductive cycle of the razor clam, solen grandis, were investigated monthly by histological and cytological observations. Samples were collected from natural intertidal population at Oshik-do, Kunsan, Korea, for one year, beginning from January to December, 1993. solen grandis is dioecious. Morphological structures of the spermatozoon of this species ar esimilar to those of other bivalve spermatozoa having a primitive type; i.e., a small head, a cap-shaped acrosome and a short mid-piece with four mitochondria surrounding axial filament. The head of spermatozoon is approximately 2 \mu m in length and sperm tail is about 20 \mu m long. The axoneme of tail flagellum consists of nine pairs of peripheral microtubules at the periphery and a pair of central microtubules at the center. Four spherical mitochondria form the paranucleus. Spawning occures once a year between early June and July, and the main spawning was observed in July when seawater temperature reaches above 20 \circ C. The reproductive cycle of male razor clam can be divieded into fivesuccessive stages; early active (December to january), late active (January to march), mature (March to early August), partially spawned (June to July), and spent/inactive stage (August to December).

• Development and Reproduction
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• v.5 no.2
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• pp.167-173
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• 2001

To characterize the difference in glycoconjugates of mouse epididymis, lectin labeling of the tissue section was conducted using Ulex europaeus agglutinin I(UEA I), succinylated wheat germ agglutinin(sWGA), and Griffonia simplicifolia lectin-I(GSL-I). UEA I which binds to outer $\alpha$-L-fucose residue that is a terminal sugar of the side chain branched from oligosaccharide chain gave the labeling in the proximal caput epithelia exclusively. Lumen was commonly labeled in all of the organ. It suggested that the glycoconjugates bearing outer $\alpha$ -L-fucose residue were largely expressed in the initial segments ot epididymis and subjected to secretion. GSL-I which binds to terminal $\alpha$ -D-galactosyl residue of glycoconjugates gave the labeling in the cytoplasm of clear cells and basal cells, and cilia in corpus and cauda regions but not in the caput region. There was no vast difference in labeling pattern by sWGA which binds to N-acetyl-glucosamine residue among the epididymal regions. Clear cells in corpus and cauda epithelia showed more intense labeling by sWGA compared to principal cells, suggesting the functional specialization of this type of cells. The labeling intensities of luminal content by UEA I and sWGA decreased in cauda region compared to corpus region suggesting the presence of enzymatic activities responsible for processing the $\alpha$-L-fucose and N-acetyl-glucosamine residues from secreted glycoconjugates. In summary, the difference in glycoconjugates bearing the $\alpha$-L-fucose, $\alpha$-D-galactose, and N-acetyl-glucosamine residues according to the type of epithelial cells and epididymal segments suggests functional specialization and different roles of each segment in the processing of sperm surface antigens during the epididymal transit.

• Development and Reproduction
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• v.2 no.1
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• pp.89-99
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• 1998

This study was carried out to investigate the morphological changes of gonadotropes in pituitary gland and spermatogenic cells in testis, obtained from 150 of 3-year-old immature and mature male rainbow trout (Oncorhynchus mykiss) during the reproductive cycles from March to February in the following year. In the maturation cycle of the pituitary gonadotropes of cultured rainbow trout, three periods can be distinguished i.e. a period of resting(March-August), a period of full spermatogenesis (September-November), and a period of breeding (December-February). The ultrastructures of the gonadotropes largely parallel the cyclical changes in the tests. The seminiferous tubules contain all spermatogenetic stages and sperm cells in a period of early maturation. At first, the size of the nucleus and cytoplasm decrease gradually at every stages from spermatogonia to spermatids. In the secondary spermatocytes, the small mitochondria are located over the outer cytoplam. In spermatids, the cytoplasmic masses move toward the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm. In spermatids, the cytoplasmic masses move towark the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm and begin to elongate. In spermatozoa, the surface of the nucleus devreases in volume. Examination by TEM shows that the nuclear envelope and plasma membrane are slightlywrinkled and closely adhered to the nucleus of spermatozoa. Two oval mitochondria are quite separated and the flagellum is inserted into the base of the spermatozoa head.The axoneme in this fish has the typical pattern such as nine peripheral doublets and a central doublet(9+2). there are remarkable individual differences in the size and morphology of spermatozoa head as observed by transmission and scanning electron microscopy.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.109-109
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• 2002

In recent reports, the multiple reproductive defects such as cryptorchidism, hypospadias, epididymal cysts, low sperm counts, and testicular cancers are increased in humans, and these changes were doubted by the chemicals with estrogenic or antiandrogenic activities in our environment. To compare the effects of neonatal exposure of di (n-butyl) phthalate and flutamide on the development of reproductive organs and to identify the specific mechanisms of these abnormalities related to the male reproducton, Sprague-Dawley neonate male rats were injected subcutaneously during 5-14 days after birth with corn oil (control), flutamide (0.05, 0.1, and 0.5 mg/animal) and DBP (5, 10, and 20 mg/animal). Animals were killed at 31 (immature) and 42 (pubertal) days of age respectively and blood was collected from abdominal aorta for serum testosterone analysis. Testes, epididymides, seminal vesicles, ventral prostate, levator ani plus bulbocavernosus muscle (LABC), cowpers glands and glans penis were weighed. Expression of steroid hormone receptors (AR and ER) was examined in the testes and ventral prostate. At 31 days of age, ventral prostate, seminal vesicles, LABC, and cowpers glands significantly decreased in the flutamide (0.5 mg/animal) and DBP (20 mg/animal), but serum testosterone levels were not changed. Flutamide slightly delayed the testes descent at the high dose (0.5 mg/animal), but DBP did not show any significant effect on the testes descent at all doses. DBP and flutamide decreased the expression of AR protein in the testes but did not affect the expression of ERa and ER protein in the testes. At 42 days of age, ventral prostate, seminal vesicles, and cowpers glands weights were still significantly decreased at the high dose of flutamide (0.5 mg/animal) and DBP (20 mg/animal), but the weights of testes and epididymides were not different. Serum testosterone decreased significantly in DBP treated animals and slightly, not significantly, in flutamide group. While DBP still significantly decreased the expression of AR protein in testis, flutamide recovered from downregulation of AR protein and did not affect the expression of ERa and ER protein in the testes. Based on these results, flutamide and DBP have shown several similar patterns in reproductive abnormalitis, but some marked differences which may be caused by different acting mechanism.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.90-90
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• 2002

The purpose of this study was to investigate the effect of kind and combination of sugars on the viability and acrosome damage of post-thaw spermatozoa in canine. The extender used was Tris-citric acid extender (Tris-buffer) supplemented with 20% Egg-yolk, 8% glycerol, 1% Equex STM paste, and 70 mM sugars such as monosaccharide (fructose and xylose) and disaccharide(trehalose). To evaluate of sugar combination, the sugars supplemented in Tris-buffer were combined such as single (fructose, xylose, trehalose), two combinations (Fruc+Tre, Fruc+xyl, Tre+xyl) and three combinations (Fruc+Tre+Xyl). The concentration of sperm collected were adjusted of 50${\times}$10$\^$6/ per straw for freezing. The frozen spermatozoa were thawed at 37$^{\circ}C$ for 1 min and then analysis for CASA program in Livestock Improvement Main Center, NACF. The motility of post-thaw spermatozoa in Fruc+Tre was higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (79％ vs. 63, 66, 70, 71, 74 and 75%). The progressive motility after CASA analysis in Fuc+Tre group was also higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (67% vs. 53, 57, 60, 61, 62 and 64%). The acrosome damage of post-thaw spermatozoa stained was not significantly different among treatment groups such as fructose, trehalose, xylose, Fru+Tre, Fru+xyl, Tre+xyl and Freu+tre+xyl (17.7, 18.3, 28.0, 17.0, 19.7, 20.0 and 19.0%). The results indicated that the motility and progressive motility of post-thaw spermatozoa in Fru+Tre group was better, and acrosome normality was not different among all groups. The use of Tris-buffer supplemented with Fru+Tre as sugar for frezing of canine spermatosoa could be better and apply to semen banking and artificial insemination.

• Korean Journal of Ichthyology
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• v.13 no.1
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• pp.74-84
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• 2001

The early developmental stages, growth and morphological changes of the Korean bullhead, Pseudobagrus fulvidraco, were studied from a series of reared specimens. Details of the early developmental stages are illustrated with special reference to morphological transformations. Egg and sperm of Korean bullhead were obtained from mature adults under hormonal treatment, fertilized artificially, and incubated in the aquarium. The incubation period of fertilized eggs was 55 to 66 hours at a temperature of 24.9${\pm}$0.34$^{\circ}$. Larvae were fed successively with Artemia salina and Daphnia magna for 2 to 15 days and artificial food after 20 days. Fertilized eggs were adhesive and spherical with a diameter of 2.04mm(n = 100). The mean total length of newly hatched larvae was about 4.92${\pm}$0.33 mm. Mouth opening occurred on one-day-old yolk-sac larvae, and initial feeding was observed on the third day after hatching. The morphological transitions from larvae to juvenile and juvenile to young stages occurred when the fish reached about 17 mm in total length (about 13days after hatching) and about 32 mm in total length (about 30 days after hatching), respectively. Many changes in proportion of body parts to total length were observed at about 7~8 mm and 30~32 mm, corresponding to the transformations from larvae to juvenile and from juvenile to young, respectively. In comparing relative growth of each body part against total length, those characteristics related to head parts showed positive growth in the pre-larval stages, while those concerning mobile abilities showed positive growth in the post-larval stage.

• Journal of Environmental Health Sciences
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• v.42 no.1
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• pp.53-60
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• 2016

Objectives: Preservatives are commonly used in pharmaceuticals, cosmetics and other products to extend the expiration date and prevent the growth of microorganisms. Preservatives are generally effective in controlling mold and inhibiting yeast growth, and against a wide range of bacterial attacks as well. They also adversely affect the quality of sperm and cause precocious puberty in children. This study was performed to analyze seven preservatives used in pharmaceuticals and personal care products. Methods: Five kinds of pharmaceuticals and personal care products (PPCPs) were examined for analysis with a high performance liquid chromatography-diode array detector. Each sample was homogenized and the targeted compounds were extracted with methanol. The suspended particulate was removed by syringe filter. Next, the sample was injected into an HPLC system. The separation of the seven preservatives was achieved with a C18 column and gradient mode. The accuracies were between 73% and 120% and precision was lower than 11.58% (RSD). Results: All of the calibration curves showed good linearity with a coefficient of determination ($r^2$) over 0.999. Among the PPCP samples, the detection rate of preservatives was 32.5% for pharmaceuticals, 44.8% for toothpaste, 76.9% for mouthwash, 40.0% for body lotion and 56.0% for wet tissues. The average concentrations of the preservatives in PPCPs were BA 1141.0 mg/kg, MP 709.8 mg/kg, EP 624.9 mg/kg, PP 216.9 mg/kg, BP 167.8 mg/kg, and TCS 538.2 mg/kg. The most frequently detected preservatives in pharmaceuticals and personal care products were BA, MP and PP. The concentrations of preservatives exceeded Korean regulatory standards in 11 samples of medicines, three of mouthwash and two of body lotion. Conclusion: We found that most of the PPCP samples contained various preservatives. It is necessary to identify which preservatives were used and to determine the level of preservatives in PPCPs and to assess the health risk to susceptible populations such as children.

• Interdisciplinary Bio Central
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• v.2 no.4
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• pp.12.1-12.7
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• 2010

The National Bio Resource Project-Rat (NBRP-Rat) comprises the largest bank of laboratory rat (Rattus norvegicus) strains in the world. Its main focus is to develop infrastructure that will facilitate the systematic collection, preservation, and provision of rat strains. To breed effectively more than 180 rat strains in living stock, we establish the genetic control system in which a systematic set of genetic diagnoses and genetic monitoring are included. Genetic monitoring is performed by using 20 polymorphic markers. Monitoring is carried out when a living animal stock is re-established by using cryopreserved embryos or sperm or when a rat strain is first introduced to the NBRP-Rat by a depositor. Additional monitoring is then carried out on each strain every two years. Genetic diagnosis is performed largely by employing the Amp-FTA method. Protocols which detail how to perform a genetic diagnosis of 11 transgenes and 24 mutations have been made. Among the mutations, nine can be detected by simple gel electrophoresis of the PCR products, 11 by restriction enzyme treatment of the PCR products, and four by direct PCR product sequencing. Using this genetic control system, the NBRP-Rat can guarantee the genetic quality of its rat strains.