• The Korean Journal of Zoology
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• v.35 no.2
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• pp.211-218
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• 1992

한국산 노랑초파리 군에 속하는 8종의 유전적 유연관계를 밝히고자 생식적 격리 그리고 수용성 단백질을 전기영동법으로 분석하였다. 생리적 격리 실험에서 교배전격리 실험결과를 Watanane와 Kawanishi model에 근거하여 보면 D. aurario complex 3종 중 D. triauror물가 원시종이며 남 auraria는 팍생종으로 나타났다. 교배후 격리 실험에서 나. melonogaster와 D. simuions의 교배시, D. melonogoster를 수컷으로 하였을 때는 불임의 수컷만이, 또 D. melonogaster를 암컷으로 곯였을 때는 불임의 암컷만이 출현하였다. 그리고 남 ouraria complex 3종간의 교배에서는 수정 능력이 있는 수컷과 암컷이 출현하였는데, 이는 아직 남 auraria Complex가 semispecies단계에 있음을 나타내는 것이라 할 수 있다. SDS-PAGE로 분석한 노랑초파리 군 8종의 band pattern을 densitometer로 scanning한 결과 남 susu상가 가장 특이하였으며, TDE에 의한 유전적 거리(Aquadro and Avise's)는 남 auror지와 봉 triauror지사이가 0.155로 가장 낮았고, D. melonogaster와 고. mfa 사이가 0.422로 가장 높았다. 본 연구의 결과를 UPGMA법 뜨로 분석하면, 한국산 노랑초파리 군 8종은 4개의 아군으로 나뉘어지며 이들은 2개의 다른 큰무리로 구분되었는데, D. suzu가기 아군, D. lutescens의 아군, D. melanogoster와 봉 simulons의 아군이 속한 큰무리와, D. mfa,0. ouroria, D. biauroria 그리고 남 triourario가 속한 다른 큰무리로 나눌 수 있다.

• Advances in Traditional Medicine
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• v.6 no.2
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• pp.86-95
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• 2006

The ethanol extract of the Crotalaria juncea seeds, which showed promising antispermatogenic and antiandrogenic activities in albino mice, was taken up further for the isolation of the active fractions present in it. Two fractions that were obtained from thin layer chromatography were subjected for testing to know their antispermatogenic and antiandrogenic activities. After preliminary trials the fraction I showed maximum antifertility activity at the dose level of 200 mg/kg body weight when administered orally to the rats for 50 days. The fraction I was found to affect spermatogenesis as well as the endocrine functions of the testis as indicated by gravimetric, histopathological and biochemical changes. Further this fraction has caused degenerative changes in the seminiferous tubules and Leydig cells of the testis. The accessory reproductive organs like epididymis, seminal vesicles, vas deferens, prostrate, Cowper's gland and Levator Ani muscle showed significant malfunction. Cauda epididymal sperm count and sperm motility were reduced significantly. The treatment has also resulted in increase in the cholesterol level and alkaline phosphatase activity, and decrease in protein, glycogen, sialic acid contents and acid phosphatase activity in testis. It is noteworthy that RIA studies have shown significant reduction in serum FSH, LH and testosterone. Scanning electron microscopic observations revealed abnormalities in sperm structure.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.163-167
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• 2006

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.3
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• pp.183-190
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• 2001

Objective: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. Methods : Mouse ovaries were fixed and cut into serial sections (7 im thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell $II^{TM}$ system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). Results: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GA PDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 $\grave{i}$ l with 50 oocytes, thus the resting 19.75 il cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. Conclusion: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.4
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• pp.199-206
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• 2016

Objective: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. Methods: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. Results: The sperm DFI showed a significant correlation (r=-0.347, p< 0.001) with sperm motility and morphology (r=-0.114, p< 0.05) but not with other semen parameters. The DFI ($11.5%{\pm}2.0%$) of semen samples was significantly reduced by DGC ($8.1%{\pm}4.1%$) or MACS alone ($7.4%{\pm}3.9%$) (p< 0.05). The DFI was significantly further reduced by a combination of DGC and MACS ($4.1%{\pm}1.3%$, p< 0.05). Moreover, the combination of DGC and MACS ($1.6%{\pm}1.1%$, p< 0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC ($4.4%{\pm}3.2%$) or MACS alone ($3.4%{\pm}2.2%$). Conclusion: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.

• Korean Journal of Plant Taxonomy
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• v.47 no.4
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• pp.289-296
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• 2017

The three self-compatible, terrestrial orchids Cephalanthera erecta, C. falcata, and C. longibracteata flower synchronously in sympatric populations. Cephalanthera erecta and C. longibracteata, which have white flowers that do not fully open, are predominantly autogamous, whereas the food-deceptive C. falcata, whose bright yellow flowers open completely, is predominantly outcrossing. The formation of hybrids rarely occurs between species owing to strong prepollination barriers (floral isolation). If these three species have evolved toward the prevention of interspecific hybridization, we can expect that the spatial distribution patterns of the three species would be characterized as spatial segregation (i.e., 'spatial repulsion') from each other. To test this prediction, we studied the three Cephalanthera species in sympatric populations showing coincident flowering within Yeonwhasan Provincial Park (YPP, Gyeongsangnam Province, South Korea). We found strong spatial aggregation in each population and spatial independence in the interspecific spatial distribution, differing from previous studies. We further hypothesize that Cephalanthera species in sympatry within YPP are distributed somewhat randomly in space, perhaps due to the sharing of similar mycorrhizal fungi.

• Korean Journal of Ichthyology
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• v.2 no.1
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• pp.88-94
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• 1990

Natural hybrids between Pungitius sisensis sinensis and P. sinensis kaibarae were found in their sympatric area, a streamlet below Chibyon Reservoir in Kangnung city, Kangwondo, Korea. P. sinensis sinensis and P. sinensis kaibarae showed the same electropherograms as reported previously. However, about 10% of the specimens had peculiar banding patterns and they seemed to be hybrid between the two subspecies. They are divided further into two groups: sinensis-type and kaibarae-type hybrid. Two morphometric characters, snout length and depth of caudal peduncle, indicated the hybrid's property well. The color of testis of P. sinensis sinensis and P. sinensis kaibarae was grey and black, respectively, but that of hybrid was intermediate. The problem of reproductive isolation between the two groups raised by the finding of their hybrids was discussed.

• Parasites, Hosts and Diseases
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• v.51 no.6
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• pp.629-632
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• 2013

Members of the genus Trichinella are small nematodes that can infect a wide range of animal hosts. However, their infectivity varies depending on the parasite and host species combination. In this study, we examined the susceptibility of 4 species of laboratory rodents, i.e., mice, rats, hamsters, and gerbils to Trichinella papuae, an emerging non-encapsulated Trichinella species. Trichinella spiralis and Trichinella pseudospiralis were also included in this study for comparison. Fifteen animals of each rodent species were infected orally with 100 muscle larvae of each Trichinella species. Intestinal worm burden was determined at day 6 and 10 post-inoculation (PI). The numbers of muscle larvae were examined at day 45 PI. The reproductive capacity index (RCI) of the 3 Trichinella species in different rodent hosts was determined. By day 6 PI, 33.2-69.6% of the inoculated larvae of the 3 Trichinella species became adult worms in the small intestines of the host animals. However, in rats, more than 96% of adult worms of all 3 Trichinella species were expelled from the gut by day 10 PI. In gerbils, only 4.8-18.1% of adult worms were expelled by day 10 PI. In accordance with the intestinal worm burden and the persistence of adults, the RCI was the highest in gerbils with values of $241.5{\pm}41.0$ for T. papuae, $432.6{\pm}48$ for T. pseudospiralis, and $528.6{\pm}20.6$ for T. spiralis. Hamsters ranked second and mice ranked third in susceptibility in terms of the RCI, Rats yielded the lowest parasite RCI for all 3 Trichinella species. Gerbils may be an alternative laboratory animal for isolation and maintenance of Trichinella spp.

• BMB Reports
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• v.45 no.2
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• pp.96-101
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• 2012

Urea-based nitrogen fertilizer was widely utilized in maize production, but transporters involved in urea uptake, translocation and cellular homeostasis have not been identified. Here, we isolated three maize aquapoin genes, ZmNIP2;1, ZmNIP2;4 and ZmTIP4;4, from a cDNA library by heterogous complementation of a urea uptake-defective yeast. ZmNIP2;1 and ZmNIP2;4 belonged to the nodulin 26-like intrinsic proteins (NIPs) localized at plasma membrane, and ZmTIP4;4 belonged to the tonoplast intrinsic protein (TIPs) at vacuolar membrane. Quantitative RT-PCR revealed that ZmNIP2;1 was expressed constitutively in various organs while ZmNIP2;4 and ZmTIP4;4 transcripts were abundant in reproductive organs and roots. Expression of ZmTIP4;4 was significantly increased in roots and expanded leaves under nitrogen starvation, while those of ZmNIP2;1 and ZmNIP2;4 remained unaffected. Functions of maize aquapoin genes in urea transport together with their distinct expression manners suggested that they might play diverse roles on urea uptake and translocation, or equilibrating urea concentration across tonoplast.

• Fisheries and Aquatic Sciences
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• v.12 no.4
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• pp.276-285
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• 2009

Despite great commercial interest, relatively little has been described about molecular mechanism of bivalve reproduction. We investigated genes involved in ovarian maturation of the Yesso scallop, Patinopecten yessoensis. GSI index and histological analysis revealed that maturation of ovary begin in February and spawning period is from April to June which is similar to the previous study in the East Sea. As result of combination analysis of differential display RTPCR (DDRT-PCR) and histological examination, vitellogenin (Vg), ferritin (Ft) and ADT/ATP carrier protein (ACC) were identified as differently expressed genes in maturating ovary. Endpoint RT-PCR results showed that Vg is ovary-specific genes whereas Ft and ACC are expressed ubiquitously suggesting that Vg can be good molecular markers for ovarian development and sex determination in bivalves. Quantitative PCR results revealed that Vg were expressed highest during growth stage and appears to play a major role in oocyte maturation. On the contrary, expression of Ft was highest after spawning stage, which suggests that up-regulation may be involved in spawning and inactive stages in which the scallops recover from spawning. In addition, high level of the mitochondrial gene, ACC, may play a role in energy metabolism in maturating oocytes. Isolation and molecular studies of these key genes will expand our knowledge of the physiological changes from various exogenous factors including temperature, salinity, pH, even or numerous endocrine disrupting chemicals (EDCs) during reproductive cycle. In addition, further study of these genes implicates various industrial applications including the stable seed production, increased food quality, or economic aquaculture system.