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Immunohistochemical study of the gastrointestinal endocrine cells in the Mongolian Gerbils, Meriones unguiculatus (몽골리안 저빌(Meriones unguiculatus)의 위장관 내분비 세포에 대한 면역조직화학적 연구)

  • Lee, Jae-hyun;Lee, Hyeung-sik;Ku, Sae-kwang;Park, Ki-dae;Kim, Kil-soo
    • Korean Journal of Veterinary Research
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    • v.40 no.4
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    • pp.653-660
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    • 2000
  • In order to study the regional distribution and relative frequency of the immunoreactive endocrine cells in the gastrointestinal tract of the Mongolian gerbil, Meriones unguiculatus, the gastrointestinal tract was divided into 9 portions (cardia, fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) and immunostained by immunohistochemical (PAP) method using 8 types of specific antisera against cholecystokinin (CCK)-8, gastrin, secretin, pancreatic polypeptide(PP), somatostatin, serotonin, glucagon and insulin. CCK-8-, gastrin-, somatostatin- and serotonin-immunoreactive cells were demonstrated in this study. These immunoreactive cells were found in the gastric gland regions of the pylorus or between parietal and chief cells of the fundus with round to spherical shaped, and in the interepithelial regions of the intestinal tract with spherical to spindle shaped except for jejunum where some of immunoreactive cells were also observed in the intestinal glands with round to spherical shaped. CCK-8-immunoreactive cells were restricted to the pylorus and duodenum with numerous and a few frequency, respectively. Gastrin-immunoreactive cells were restricted to the pylorus with numerous frequency. Similar to those of gastrin-immunoreactive cells, somatostatin-immunoreactive cells were restricted to pylorus with moderate frequency. Serotonin-immunoreactive cells were detected throughout whole gastrointestinal tract except for cardia and cecum with moderate or numerous frequency. However, no secretin-, PP-, glucagon- and insulin-immunoreactive cells were observed in this study. From these results, the appearance type, regional distribution and relative frequency of immunoreactive endocrine cells in the gastrointestinal tract of the Mongolian gerbils were somewhat lowered or restricted compared to those of other mammals and these differences were might be caused by feeding habits and species specification.

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Dynamin II Expression and Morphological Comparison of NIH3T3 and NIH3T3 (ras) Cells (NIH3T3와 NIH3T3(ras) 세포에서 Dynamin II 발현 및 형태적 비교)

  • Lee, Chul-Woo;Kim, Su-Gwan;Choi, Jeong-Yun;Choi, Baik-Dong;Bae, Chun-Sik;Jeong, Soon-Jeong;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.35 no.3
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    • pp.121-128
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    • 2005
  • It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.

Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells (미성숙 매복지치의 치낭, 치수, 치근유두 조직에서 다능성 줄기세포의 분리와 특성화에 대한 연구)

  • Song, Jung-Ho;Park, Bong-Wook;Byun, June-Ho;Kang, Eun-Ju;Rho, Gyu-Jin;Shin, Sang-Hun;Kim, Uk-Kyu;Kim, Jong-Ryoul
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.3
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    • pp.186-196
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    • 2010
  • Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

Comparative Histological Study on the Parafollicular Cells of Mammals (각종 포유동물 갑상선내의 소포방세포에 관한 비교조직학적 연구)

  • 고정식;박상윤
    • The Korean Journal of Zoology
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    • v.23 no.2
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    • pp.89-108
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    • 1980
  • Comparative studies on the parafollicular cells of the some mammalian species from five different orders were carried out; i.e., man from Primates, cattle, pig, and black goat from Artiodactyla, dog from Carnivora, rabbit from Lagomorpha, rat, mouse, and squirrel from Rodentia. For this study, various special techniques for the parafollicular cells, including Grimelius' silver impregnation method (Sawicki and Bajko, 1974), Singh's argentaffin method (Singh, 1964), HCl-toluidine blue stain (Sawicki, 1971), and HCl-lead hematoxylin stain (Solcia et al., 1969), were applied. Authors obtained the following results: 1. Number of parafollicular cells in the same area of thyroid tissues are significantly different from species to species. Number of cells were largest in dog and less cells were found in the following orders; rat, squirrel, mouse, rabbit, cattle, pig, black goat and finally the smallest number in man. 2. Distribution of parafollicular cells within thyroid gland are significantly different from portion to portion in case of cattle, rabbit, squirrel and mouse, but it is not significant in dog, man, pig, black goat and rat (see Table 1-1 and 1-2). 3. In dog, clustered parafollicular cells are located usually in the interfollicular space, and groups of parafollicular cells are located in the para-and/or inter-follicular positions in rabbit. But in the other animals parafollicular cells are found solitarily in the intra-and/or para-follicular positions. 4. The shape of parafollicular cells shows oval to round contour in dog, but it is polymorphic, for example, spindle, conical, oval, round or elongated with cytoplasmic processes, in the other animals. 5. Size of parafollicular cells is larger in cattle, dog and pig, smaller in rat, mouse and squirrel, and medium-size in rabbit, man and black goat. 6. Parafollicular cells of pig, cattle, dog and squirrel are observed to contain densely packed granules, whereas those of mouse, rat and man contain relatively scanty granules. 7. Parafollicular cells of all the mammals show more or less positive reaction to Grimelius' argyrophile silver impregnation method, HCl-toluidine blue stain and HCl-lead hematoxylin stain, whereas they show negative reaction to argentaffin method (see Table 2). 8. Considering the above finding, it is concluded that there are species differences in the distribution, location and shape of parafollicular cells, and infer that preferable staining method should be selected for reliable detection of parafollicular cells, beacuse staining methods applied on the cells in this study show variable reactions according to species.

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Phospholipase C-γ Activation by Direct Interaction with β-Tubulin Isotypes (베타 튜불린에 의한 포스포리파제 C-감마1의 활성화)

  • Lee, In-Bum;Kim, Sung-Kuk;Choi, Jang-Hyun;Suh, Pann-Ghill;Chang, Jong-Soo
    • Journal of Life Science
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    • v.16 no.4
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    • pp.612-617
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    • 2006
  • Phosphoinositide-specific phospholipase $C-{\gamma}\;1\; (PLC-{\gamma}\;1)$ has pivotal roles in cellular signaling by producing second messengers, inositol 1,4,5-trisphosphate $(IP_3)$ and diacylglycerol (DG). Tubulin is a main component of microtubules and mitotic spindle fibers, which are composed of ${\alpha}-$ and ${\beta}-tubulin$ heterodimers in all eukaryotic cells. In humans, six ${\beta}-tubulin$ isotypes have been identified which display a distinct pattern of tissue expression. Previously we found that $PLC-{\gamma}\;1$ and one of four ${\beta}-tubulin$ isotypes including ${\beta}1$, ${\beta}2$, ${\beta}3$ and ${\beta}6$, colocalized in COS-7 cells and cotranslocated to the plasma membrane to activate $PLC-{\gamma}\;1$ upon agonist stimulation. In the present study, we demonstrate that the remaining two, tubulin ${\beta}4$ and ${\beta}5$, also showed a potential to activate $PLC-{\gamma}\;1$. The phosphatidylinositol 4,5-bisphosphate $(PIP_2)$ hydrolyzing activity of $PLC-{\gamma}\;1$ was substantially increased in the presence of purified ${\beta}4$ and ${\beta}5$ tubulin in vitro, whereas the activity was not promoted by bovine serum albumin, suggesting that tubulin ${\beta}4$ and ${\beta}5$ also activate $PLC-{\gamma}\;1$. Taken together, our results suggest that all the ${\beta}-tubulin$ isotype activates $PLC-{\gamma}\;1$ activity to regulate cellular signaling.

Electron Microscopic Studies on Olfactory Bulbs in the Vertebrates by Phylogenetics (계통발생에 따른 척추동물의 뇌후구에 대한 전자현미경적 연구)

  • Choi, W.B.;Chung, Y.H.;Seo, J.E.
    • Applied Microscopy
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    • v.15 no.2
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    • pp.31-68
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    • 1985
  • Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.

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Formation of Metastases in C57 BL/6 Mouse Lung (LL/2 Cells접종에 의한 C57 BL/6 Mouse 폐에서의 전이과정)

  • Kim, Ji-Hyun;Lee, Ji-Young;Kim, Woo-Kap;Hwang, Woo-Ik
    • Applied Microscopy
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    • v.29 no.2
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    • pp.137-147
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    • 1999
  • Comparative differences between the fine structure of cultured LL/2 cell in vitro and tumor cells in vivo which were induced in the lung by inoculation of LL/2 cells to C57 BL/6 mouse via tail vein during 21 days are not observed except for cell configuration which was changed spindle shape into oval shape. At first tumor cells appeared at lymphatic nodules and around capillary in the lung. Tumor cells divided actively by mitosis, so they became tumor nodules. The pulmonary aveoli around tumor nodules were observed somewhat flattened in shape but the cells in the aveoli appeared to be in normal condition. Furthermore the normal lung cells were observed in the tumor nodules and some apoptotic tumor cells appeared in the large tumor nodules. A lot of neutropiles were observed in the aveoli and tumor nodules of C57 BL/6 mouse lung after inoculation 22 days and 31days.

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Evaluation of Early Productivity of High Density 'Fuji' Apple Orchards by Planting Well-feathered Trees/M.9 EMLA ('후지' 사과의 우량 측지묘/M.9 EMLA를 이용한 밀식재배원의 조기 생산성 평가)

  • Yang, Sang-Jin;Park, Moo-Yong;Song, Yang-Yik;SaGong, Dong-Hoon;Yoon, Tae-Myung
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.374-380
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    • 2010
  • Well-feathered (5.2 feathers, stem diameter 13 mm) trees of 'Fuji' apple/ M.9 EMLA were planted at $4.0{\tiems}1.5$ m and whip trees (stem diameter 10 mm) of 'Fuji'/M.26 at $4.0{\times}2.0$ m were trained to the slender spindle. The productivity and yield efficiency of two orchard systems were compared for 6 years. The canopy volume of the tree/M.9 EMLA reached $2.07m^3$ in 2nd year and increased slowly to almost the targeted tree volume of $2.9m^3$ in 4th year. Trees/M.26 grew slowly at the begin but from 3rd year the tree volume expanded quickly to reach $5.6m^3$ in 5th year, covering over the allowed space. Yield of M.9 EMLA per 10a increased from 0.3 ton in 2nd year to 4.6 ton in 5th year, and 5.0 ton in 6th year, but yield of M.26 per 10a increased from 0.5 ton in 3rd to 2.9 ton in 6th year. Cumulative yield per 10a up to 6th year was 13.9 ton for M.9 EMLA but only 9.8 ton for M.26. Fruit weight for M.9 EMLA was heavier than that for M.26. In conclusion, the high density planting system with well-feathered trees/M.9 EMLA was better than the conventional wide planting system with whip trees/M.26.

The Characteristics of Dolmen Culture and Related Patterns during the End Phase in the Gyeongju Region (경주 지역 지석묘 문화의 특징과 종말기의 양상)

  • Lee, Soohong
    • Korean Journal of Heritage: History & Science
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    • v.53 no.4
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    • pp.216-233
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    • 2020
  • This study set out to review tomb culture in the Gyeongju region during the Bronze Age, and also examine the patterns of dolmens during their end phase. For these purposes, the study analyzed 18 tomb relics from the Bronze Age and nine from the early Iron Age. Gyeongju belongs to the Geomdan-ri cultural zone. Approximately 120 tombs from the Bronze Age have been excavated in the Gyeongju region. There are fewer tombs than dwellings in the region, which is a general characteristic of the Geomdan-ri cultural zone. Although the number of tombs is small, the detailed structure of the dead body is varied. During the Bronze Age, tombs in the Gyeongju region were characterized by more prolific construction of pit tombs, dolmens with boundaries, and stacked stone altars than were the cases in other areas. There is a great possibility that the pit tombs in the Gyeongju region were influenced by their counterparts in the northeastern parts of North Korea, given the spindle whorl artifacts buried at the Dongsan-ri sites. Dolmens with boundaries and stacked stone altars are usually distributed in the Songguk-ri cultural zone, and it is peculiar that instances of these are found in large numbers in the Gyeongju region as part of the Geomdanri cultural zone. Even in the early Iron Age, the building of dolmens with boundaries and stacked stone altars continued in the Gyeongju region under the influence of the Bronze Age. A new group of people moved into the area, and they crafted ring-rimmed pottery and built wooden coffin tombs. In the early Iron Age, new rituals performed in high places also appeared, and were likely to provide venues for memorial services for heavenly gods in town-center areas. The Hwacheon-ri Mt. 251-1 relic and the Jukdong-ri relic are ruins that exhibit the aspect of rituals performed in high places well. In these rituals performed in high places, a stacked stone altar was built with the same form as the dolmens with boundaries, and a similar rock to the cover stone of a dolmen was used. People continued to build and use dolmens with boundaries and stacked stone altars while sustaining the Bronze Age traditions, even into the early Iron Age, because the authority of dolmens was maintained. Some dolmens with boundaries and stacked stone altars, known as being Bronze Age in origin, would have continued to be used in ritual practices until the early Iron Age. Entering the latter half of the second century B.C., wooden coffin tombs began to propagate. This was the time when the southern provinces, including the Gyeongju region, were included in the East Asian network, with the spread of ironware culture and the arrival of artifacts from central China. Around this time, dolmen culture faded into history with a new era beginning in its place.

Characterization of Traits Related to Grain Shape in Korean Rice Varieties (국내 육성 벼 품종 입형 관련 특성 분석)

  • Lee, Chang-Min;Lee, Keon-Mi;Baek, Man-Kee;Kim, Woo-Jae;Suh, Jung-Pil;Jeong, Oh-Young;Cho, Young-Chan;Park, Hyun-Su;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.199-213
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    • 2020
  • Grain size and shape are the two important components contributing to rice yield and quality. To analyze traits related to grain-shape, a total of 272 varieties derived from japonica, japonica black and Tongil-type rice accession in Korea were evaluated in this study. The traits, grain length (GL), grain width (GW), grain thickness (GT), length to width ratio (RLW), and 1000-grain weight (TGW) were measured and replicated 10 times. Genes (GW2, GS3, qGL3, qSW5, GS5, TGW6, GW7, and GW8) related to grain-shape were validated in the accessions using specific DNA marker sets. K-mean clustering of the accession based on phenotypic data revealed three groups: group 1 was classified by GW and GT and included most of japonica type, group 2 was classified by RLW and GL reached a medium size and possessed a half spindle-shaped type, and group 3 was classified by TGW, reached a long size and possessed a semi-round shape. In validation tests using the marker sets, both gw2 and tgw6 were validated in less than 1% of the tested accessions and two allelic types, qgl3 and gw8, were only verified in Tongil-type accessions. For GW8 and GW2, any different amplicons were not amplified in any japonica or Tongil-type accessions, respectively. In order to suggest the representative grain-shape gene combinations for each ecotype, the allelic combinations were evaluated by PCR analysis. Cj1 and 2 in japonica (Cj1-7), Cj_b1 and 2 in japonica-black (Cj_b1-3), and CT3 in Tongil-type (CT1-13) turned out to be the dominant combination in each ecotype, respectively. In addition, the results revealed that introgression of four genes (gw2, gs3, qSW5, and GS5) would expand the diversity of grain shape in Korean japonica varieties. The gene combinations information could be utilized practically to understand or enhance grain shape in japonica rice breeding program.