• ALGAE
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• 제24권1호
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• pp.39-45
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• 2009

The conjugation processes of a filamentous freshwater green alga Spirogyra varians were examined using FITC-lectins. Conjugation comprised five steps: 1) aligning with adjacent filaments, 2) formation of conjugation protru-sion (papilla), 3) fusion of the protrusions, 4) formation of conjugation tube,and 5) formation of zygotes. Three lectins, ConA, RCA and UEA, showed considerable labeling during the progression of conjuation. FITC-ConA labeled the surfaces of filaments throughout the whole conjugation processes. FITC-RCA labeling was observed at the conjugation protrusions only after the papilla formation. Strong labeling continued until formationg of zygotes at the contacting area where the conjugation tube developed, but no labeling was detected on the surface of vegetative filaments. The labeling decreased gradually over time and disappeared when zygotes were formed. FITC-UEA showed similar labeling pattern with FITC-RCA except that weak labeling remained after zygote formation. Inhibition experiments using RCA, UEA which are complementary to sugars L-fucose and D-galactose, showed considerable decrease of conjugation (<32% vs. 70% in control). These results suggested that the lectin-carbohydrate recognition system might be involved in the conjugation of spirogyra varians.

• Fisheries and Aquatic Sciences
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• 제1권2호
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• pp.250-254
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• 1998

Harmful micro algae isolated from Korean coastal waters, were tested with FITC-conjugated lectins and observed by epifluorescent microscopy to distinguish each other. Strain-specific sugar composition at the cell surface was suggested by the affinity of lectins to different microalgae. The micro algae Cochlodinium polykrikoides (CP-1) and Gymnodinium $A_3\;(GA_{3-1}\;1)$, are morphologically similar, but exhibited different binding activity with the lectins ECA, HPA and WGA. In Peridiniales, the micro alga Alexandrium tamarense (AT) bound HPA and WGA, but Scrippsiella trochoidea (ST-1) did not bind those lectins. Three species of Prorocentrum also exhibited different binding specificity with HPA, PHA and SBA. A non­toxic Korean isolate of Heterosigma akashiwo (HA-2) bound ConA, PEA and UEA. These results suggest that lectins are useful in discriminating morphologically similar species, as well as different species or strains within the same genus.

• Clinical and Experimental Reproductive Medicine
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• 제29권3호
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• pp.179-186
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• 2002

Objective: Lectins are cell-agglutinating and sugar specific proteins or glycoproteins of non-immune origin that precipitate glycoconjugates having saccharides of appropriate complementarity. Because of these properties, plant lectins have been used to help characterize the carbohydrate moieties of glycoproteins in the zona pellucida (ZP) of several mammalian species including pigs. Treatment of oocytes with various lectins blocks sperm binding to the ZP in various mammalian species. This study was undertaken to examine the distribution of sugar residues in the ZP of pig oocytes matured in vitro and the ability of spermatozoa to bind to ZP and in vitro penetration in oocytes treated with fluorescein isothiocyanate (FITC)-labelled lectins. Materials and Methods: The lectins of Banderiaea simplicifolia (BS-II, bind to $\beta$-D-N-acetylglucosamine), Canavalin ensiformis (Con A, bind to $\alpha$-D-Mannose), Lens culinaris (LCA, bind to a-D-Mannose), Ricinus communis (RCA-I, bind to $\beta$-D-Galactose) and Ulex europaeus (UEA-I, bind to $\alpha$-L-Fucose) were examined for spermatozoa penetration, binding capacity to ZP and distribution of lectins. Results: The penetration rates were significantry (p<0.05) higher in control oocytes (63%) than those treated with all lectins, but penetration rates ($40{\sim}49%$) were simililar in group treated with lectins. The incidence of monospermy was similar in oocytes untreated and UEA-I, but it was higher in oocytes treated with BS-II, Con A, RCA-I and LCA. The porcine oocytes cultured for 48 h in TC-199 medium were freed from cumulus cells and treated for 30 min with fluorescein isothiocyanate-labelled lectins. When examined under fluorescein illumination, higher (p<0.001) proportions of oocytes showed fluorescein of zona pellucida after treatment with Con A (93%), LCA (93%) and RCA-I (100%) than BS-II (37%) and UEA-I (50%). All of the oocytes treated with RCA-I exhibited strong fluorescein in the outer region of the zona pellucida while those treated with LCA exhibited strong fluorescein throughout the zona pellucida. BS-II bounded mainly to the outer region and UEA-I bounded mainly to the inner region of the zona pellucida, with either strong or weak fluorescein. At 120 min after insemination in vitro, fewer spermatozoa were bound to the zona pellucida of the oocytes treated with BS-II, Con-A and RCA-I. Of the lectins, Con A most inhibited sperm binding. Conclusions: These results suggest that $\beta$-D-Galactose residues in the porcine zona pellucida may act as primary sperm receptors and inducers of the sperm acrosome reaction and these sugar residues may be involved in the block to polyspermy.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2001년도 제18차 정기총회 및 학술발표 PROCEEDINGS
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• pp.69-70
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• 2001

Lectins have great potential as to determine the alternation of the distribution of cell surface carbohydrates during cellular development and differentiation. Here, we investigated the presence and distribution of cell surface carbohydrates on chicken primordial germ cells (PGCs) during the migration and gonadal stages using a variety of lectins. A total of six FITC-labelled lectins from several specificity classes were used: ConA (glucose/mannose), WGA (N-acetylglucosamine), STA (N-acetylglucosamine), DBA (N-acetylgalactosamine/galactose), UEA-I (fucose) and PHA-E (oilgosaccharide). As a results, PGC-specific binding was observed in STA. PGCs of migration stage (2.5- and 5.5-day embyos) were STA-positive whereas PGCs of 10-day embryonic gonad were not. The results suggest that N-acetylglucosamine residuse are present specifically in migrating chicken PGCs and changes during development.

• 한국가축번식학회지
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• 제17권4호
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• pp.339-346
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• 1994

정자의 구조와 기능을 이해하기 위해서는 정자세포구성분의 분리가 필요하다. 본 실험은 동결융해된 한우정액에 다양한 물리·화학적 처리를 하여 효과적인 정자세포구성분의 분리방법을 확립하고자 실시하였다. 물리적 분리방법으로는 vortexing 처리, 26 gauge needle 또는 strained 26 gauge needle을 1ml 주사기에 부착시킨 후 반복된 pumping, 동결보존액없이 반복된 동결융해처리등을 시행하였다. 또한, 화학적인 분리를 위해 trypsin, dithiothreitol, sodium dodecylsulfate, mercaptoethanol 등을 사용하였다. 모든 처리구중에서 가장 효과적인 정자두부와 미부의 절단을 strained 26 gauge needle이 부착된 주사기를 사용한 반복된 pumping에 의해 얻어졌다. 이러한 처리에 의해 95∼100%의 높은 정자구성분의 분리가 이루어졌다. 분리된 정자구성분의 두부표면의 보존여부를 알아보기 위해 250g/ml FITC-UEA 1 염색을 실시하였지만 특별한 두부표면변화는 관찰되지 않았다. 다른 물리적 처리방법들도 높은 정자구성분의 분리결과를 보여주었지만, strained 26 gauge needle를 사용한 방법에 비해서는 분리효율, 시간등 여러면에서 비효율적이었다. 화학적 처리에 의한 정자구성분의 분리결과는 물리적 처리에 비해 효과적이지 못했다. 분리된 정자두부와 미부를 각각 회수하기 위해 sucrose 용액을 2M, 1M, 0.5M, 0.25M 순으로 시험관에 넣은 후 1,000rpm에서 15분간 원심분리한 결과, 1M과 2M의 경계부분에 형성된 정자두부층을 얻을 수 있었다. 정자구성분의 효과적이고 간편한 분리방법이 본 실험에 의해 확립되어졌으며, 위의 방법에 의해 분리, 회수된 정자구성분은 생화학적 연구, 난자활성화기작의 이해등 다양한 응용연구의 기초자료로서 이용될 수 있을 것이다.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.381-381
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• 2000

Lectin binding assay was conducted on 3 A. tamarense isolates (AT-A, AT-2 and AT-6). Fatty acid composition of all 3 isolates was analyzed, and total carotenoid content and $\beta$-carotene were also determined. AT-A and AT-2 treated with different lectins in this study showed the positive response, whereas potentially toxic AT-6 did not bind DBA lectin, regardless of different growth phase, but conjugated ConA, PNA, RCA, SBA, UEA and WGA. It is possible that DBA is a desirable method for rapid and easy discrimination of highly toxic A. tamarense. AT-A, AT-2 and AT-6 comprised saturated fatty acids (49.0-61.9%), monounsaturated fatty acids (8.0-20.5%) and polyunsaturated fatty acids (23.2-30.5%). In particular, 22:6 (n-3) polyunsaturated fatty acid in AT-6 had a high abundance, compared with AT-A and AT-2. However, carotenoid content and $\beta$-carotene were not contributed to discriminate each isolate. Due to variability in biochemical composition at different isolates, possibly DBA and 22:6 (n-3) polyunsaturate fatty acid provide a good information for discrimination of AT-6.