• Clinical and Experimental Reproductive Medicine
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• 제22권3호
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• pp.253-258
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• 1995

Microtubules and microfilaments are major cytoskeletal components in mammalian ova that provide the framework for chromosomal movement and cellular division. Extensive changes of cytoskeletal organization occur during maturation and fertilization. The changes in cytoskeletons are essential for the normal meiotic maturation and for the formation of the biparental diploid genome of the embryo, and thus are repeated at each cell cycle during embryonic development. Disturbance of the cytoskeletal organization could result in abnormal gamete development and early embryonic death.

• 대한치과교정학회지
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• 제28권6호
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• pp.915-926
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• 1998

Cytoskeleton은 세포핵과 세포외 기질을 연결하고 있어서 기질에 가해지는 물리적 힘에 의해 cytoskeletal change가 유도되고 이에 의해 세포의 개조활성이 영향을 받는다고 생각되어 왔다. 본 연구는 골모세포 활성에 대한cytoskeletal change의 역할을 규명하기 위한 것으로서, 신생 백서로부터 조골세포양 세포를 분리, 배양하고 네가지 농도의 cytochalasin B(CB) 또는 colchicine(COL)을 3시간 처리하였다. 다시 배양액을 교환하고 24시간 동안 배양하여 prostaglandin $E_2\;(PGE_2)$, interleukin-6(IL-6), tumor necrosis factor-$\alpha$(TNF-$\alpha$) 및 matrix metalloproteinase-1 (MMP-1) 생산을 측정하고 통계적으로 비교하였으며 cytoskeletal protein actin 변화를 관찰하기위하여 면역형광염색하고 형광현미경으로 관찰하여 다음과 같은 결과를 얻었다: 1. CB 처리군에서 $PGE_2$ 생산이 증가되는 경향을 보였고 COL 처리군에서는 약물농도에 비례하여 증가하였다. 2. IL-6 생산은 CB농도 1.0 ${\mu}g/ml$일때를 제외하고 증가되었다. 3. TNF-$\alpha$도 CB 농도가 1.0 ${\mu}g/ml$ 일때를 제외하고 증가하였다. 4. MMP-1 생산은 CB 처리군에서 감소하는 경향을 보이고 COL 처리군에서는 변화되지 않았다. 5. CB처리군에서는 cytoskeletal actin stress fibers가 사라지고 세포모양이 둥글어지는 경향을 보였다. 이상의 결과로 미루어 보아 cytoskeletal rearrangement는 골모세포유사세포의 활성, 특히 $PGE_2$, IL-6, 및 TNF-$\alpha$같은 paracrine/autocrine factor의 생산과 관련있는 것으로 보인다.

• ALGAE
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• 제25권4호
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• pp.197-204
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• 2010

Cytoskeletal changes were observed during cell division of the green alga Zygnema cruciatum using flourescein isothiocynate (FITC)-conjugated phallacidin for F-actin staining and FITC-anti-$\alpha$-tubulin for microtubule staining. Z. cruciatum was uninucleate with two star-shaped chloroplasts. Nuclear division and cell plate formation occurred prior to chloroplast division. Actin filaments appeared on the chromosome and nuclear surface during prophase, and the F-actin ring appeared as the cleavage furrow developed. FITC-phallacidin revealed that actin filaments were attached to the chromosomes during metaphase. The F-actin ring disappeared at late metaphase. At telophase, FITC-phallacidin staining of actin filaments disappeared. FITC-anti-$\alpha$-tubulin staining revealed that microtubules were arranged beneath the protoplasm during interphase and then localized on the nuclear region at prophase, and that the mitotic spindle was formed during metaphase. The microtubules appeared between dividing chloroplasts. The results indicate that a coordination of actin filaments and microtubules might be necessary for nuclear division and chromosome movement in Z. cruciatum.

• BMB Reports
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• 제29권5호
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• pp.405-410
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• 1996

Previous studies have shown that transforming growth factor beta ($TGF-{\beta}$) is a potent regulator of cell growth and differentiation. To study the effects of $TGF-{\beta}$ on cell morphology and cytoskeleton reorganization, we conducted a survey using Mv1Lu mink lung epithelial cells with antibodies to cytoskeletal proteins and an extracellular matrix protein. While the untreated cells showed a cuboidal shape of typical epithelia, the Mv1Lu cells displayed a drastic shape change in the presence of $TGF-{\beta}$. This alteration was most prominent when near-confluent cells were treated with $TGF-{\beta}$. Since the morphology alteration is known to be accompanied by the reorganization of cytoskeletal proteins in other cell types, we investigated the intracellular distribution of the three major cytoskeletal structures: microfilaments, microtubules, and intermediate filaments. In the microfilament system, $TGF-{\beta}$ induced new stress fiber formation, which was caused primarily by the polymerization of cytoplasmic G-actin. However, $TGF-{\beta}$ appeared not to induce any significant changes in microtubular structures and vimentin filaments as determined by indirect fluorescence microscopy. Finally we confirmed the rapid accumulation of fibronectin by immunoblot analysis and chased the protein locations by immunofluorescence microscopy.

• BMB Reports
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• 제29권5호
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• pp.474-480
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• 1996

We present a study of evolutionary changes in expression of actin genes among closely related sea urchin species that exhibit different modes of early development. For this purpose, polyclonal antisera raised against peptides from the carboxyl terminus of the HeCyI cytoskeletal actin of Heliocidaris erythrogramma were used. H. erythrogramma is a direct developing sea urchin that proceeds from embryonic to adult stages without an intervening feeding larval stage. Expression patterns of the CyI actin isoform were compared with those of Heliocidaris tuberculata and to a related sea urchin Strongylocentrotus purpuratus, which both produce a feeding pluteus larval stage. The CyI actin of all three species is expressed in the same cell types. However, its expression patterns have been changed with reorganization of early cell lineage differentiation, which is apparent among the three species. Thus. evolutionary changes in CyI actin gene expression patterns are correlated with not only phylogenetic relationship, but developmental mode. The implication of this observation is that evolutionary changes in expression patterns of histospecific genes may underlie the emergence of novel developmental processes.

• 생명과학회지
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• 제22권1호
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• pp.16-24
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• 2012

Polyamine은 모든 세포의 성장과 분화에 필수적인 요소지만 그 기작은 아직 정확하게 밝혀져 있지 않다. 본 논문에서는 MCF-7세포에서 spm의 세포독성 기작을 연구하였다. MTT assay 결과 저농도의 spm (<10 ${\mu}M$) 처리시 cell viability가 증가하는 반면 고농도의 spm 처리시 처리 시간과 처리 농도에 의존적으로 감소되었으며, 이는 고농도의 spm이 MCF-7 cell에 cytotoxic한 효과를 가지고 있는 것으로 사료된다. Cell cycle 분석 결과 spm 농도에 의존적으로 sub-G1 단계의 세포 양이 증가하는 것으로 나타났으며, 이는 spm이 세포분열을 억제함으로써 세포사를 유발하는 것으로 생각된다. 또한 고농도의 spm 처리 후 2시간이 지나자 cell 표면이 움츠려 들며 rounding되기 시작하여 하루가 지난 후 거의 모든 cell이 culture dish 에서 떨어진 것을 관찰할 수 있었다. 이는 spm이 세포부착에 관여하여 세포사를 유발하는 것이라 생각되며, 세포부착을 조절하는 Integrin ${\beta}1$은 저농도의 spm에선 별다른 차이를 보이지 않다가 농도가 높아질수록 조금씩 감소하였으며, cytoskeletal protein인 actin은 농도의존적으로 감소하였다. 반면 adhesion protein인 FAK는 저농도의 spm 처리시에도 급격히 감소하였다. 이는 spm이 adhesion 및 cytoskeletal proteins의 발현을 억제하는 것으로 보이며, 특히 Integrin ${\beta}1$과 actin에 비해 FAK에 더 많은 영향을 끼치는 것으로 사료된다. 단백질들의 세포막상의 분포에 관한 연구에서, membrane 상에 위치하던 Integrin ${\beta}1$은 10 ${\mu}M$의 spm 처리시 세포 내로 약간의 위치변동이 일어났으나 그 양에는 크게 차이가 없었으며, 반면에 actin은 위치상엔 큰 변화가 일어나지는 않았지만 농도에 따라 크게 감소하는 것으로 나타났다. 세포이동과 형태조절에서 중추적인 역할을 하는 FAK는 세포막 안쪽에 위치하고 있다가 spm 처리시 세포 가운데로 이동하는 모습이 관찰되었으며 그 양도 크게 감소하였다. 이상의 실험에서 세포 내 spm의 변화는 MCF-7 cell의 adhesion protein인 Integrin ${\beta}1$과 FAK, 그리고 cytoskeletal protein인 actin의 발현을 조절하여 cell attachment를 억제함으로써 세포분열과 생장을 억제하는 것으로 분석된다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1695-1701
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• 2002

The purposes of this study were to optimize the in vitro maturation (IVM) and culture (IVC) systems of rabbit oocytes. Cytoskeletal structures in the germinal vesicle stage (GV) and during IVM are also investigated. Ovaries were transported from local slaughterhouses and the cumulus-oocyte complexes (COCs) were collected from ovarian follicles (${\geq}1mm$). COCs were randomly allocated to TCM199-based medium ($T_1$, TCM-199) supplemented with $NaHCO_3$, glucose, sodium pyruvate and FSH ($T_2$), $T_2+E_2+LH$ ($T_3$), $T_3+FBS$ ($T_4$), or $T_1+E_2+LH+FSH+FBS$ ($T_5$), for IVM. In Experiment 1, COCs were retrieved from the follicles and 51 GV oocytes were fixed in the fixative (MTSB-XF) for nuclear and cytoplasmic examinations. In Experiment 2, progressive changes of both the nucleus and the cytoskeleton were examined at 0, 6, 16, and 20 h after IVM. Maturation (MR) and developmental rates were assessed in Experiment 3. Cytoplasmic microtubules (MT) were clearly observed in rabbit GV oocytes. To our knowledge, this is the first report that describes the appearance of MT structures in the GV stage ooplasm. Tremendous variations in cytoskeletal alterations were observed among treatments with the exception of the vitelline ring (VR), which is constantly visible and unchanged during maturation. Germinal vesicle breakdown (GVBD) does not occur at 6 h after onset of maturation culture. When the oocytes for IVM were collected within 2 h, results from Experiment 3 showed that rates of nuclear maturation were 42, 8, 42, 37 and 65% at 16 h of IVM for $T_1$ through $T_5$, respectively, in which $T_1$, $T_4$ and $T_5$ had significantly greater MR than those in other groups (p<0.05). Morula/blastocyst development after parthenogenetic activation ranged from 20 to 63% with significantly greater rates in $T_3$, $T_4$ and $T_5$ (p<0.05). These results suggested that oocytes recovered from slaughterhouse ovaries can be matured and parthenogenetically activated in vitro, but the MR remained low in this study. Addition of $E_2$ and LH in the medium may be beneficial for cytoplasmic maturation, but FBS exerts a nega- tive role in the subsequent development of parthenogenetic embryos when energy substrates are provided in the IVC media. More studies are required for improving the MR and further development of the GV stage rabbit oocytes.

• BMB Reports
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• 제50권1호
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• pp.5-11
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• 2017

One of the characteristics of the neurons that distinguishes them from other cells is their complex and polarized structure consisting of dendrites, cell body, and axon. The complexity and diversity of dendrites are particularly well recognized, and accumulating evidences suggest that the alterations in the dendrite structure are associated with many neurodegenerative diseases. Given the importance of the proper dendritic structures for neuronal functions, the dendrite pathology appears to have crucial contribution to the pathogenesis of neurodegenerative diseases. Nonetheless, the cellular and molecular basis of dendritic changes in the neurodegenerative diseases remains largely elusive. Previous studies in normal condition have revealed that several cellular components, such as local cytoskeletal structures and organelles located locally in dendrites, play crucial roles in dendrite growth. By reviewing what has been unveiled to date regarding dendrite growth in terms of these local cellular components, we aim to provide an insight to categorize the potential cellular basis that can be applied to the dendrite pathology manifested in many neurodegenerative diseases.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.201-208
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• 2013

Objective: To investigate changes in the invasive capacity of gastric cancer cells in vitro after expression inhibition of T lymphoma invasion and metastasis inducing factor 1 (Tiam 1) and underlying mechanisms. Methods: Using adhesion selection, two subpopulations with high ($M_H$) or low ($M_L$) invasive capacity were separated from the human gastric cancer cell line MKN-45 ($M_0$). Tiam 1 antisense oligodeoxynucleotide (ASODN) was transfected into $M_H$ cells with liposomes, and expression of Tiam 1 mRNA and protein was determined by RT-PCR and quantitative cellular-ELISA. Changes in the cytoskeleton, invasive capacity in vitro and expression of ras-related $C_3$ botulinum toxin substrate 1 (Rac 1), integrin ${\beta}1$ and matrix metalloproteinase 2 (MMP 2) between Tiam 1 ASODN transfected $M_H$ cells and non-transfected cells were observed by HE staining, cytoskeletal protein staining, scanning electron microscopy, Boyden chamber tests and cyto-immunohistochemistry. Results: A positive correlation existed between the expression level of Tiam l mRNA or protein and the invasion capacity of gastric cancer cells. After ASODN treatment ($0.43{\mu}M$ for 48 h), Tiam 1 mRNA transcription and protein expression in $M_H$ cells were decreased by 80% and 24% respectively (P < 0.05), compared with untreated controls, while invasive capacity in vitro was suppressed by 60% (P < 0.05). Morphologic and ultrastructural observation also showed that ASODN-treated $M_H$ cells exhibited smooth surfaces with obviously reduced filopodia and microspikes, which resembled $M_0$ and $M_L$ cells. Additionally, cytoskeletal distribution dramatically altered from disorder to regularity with reduced long filament-like structure, projections, pseudopodia on cell surface, and with decreased acitn-bodies in cytoplasm. After Tiam 1 ASODN treatment, the expression of Rac 1 and Integrin ${\beta}1$ in $M_H$ cells was not affected (P > 0.05), but that of MMP 2 in $M_H$ cells was significantly inhibited compared with untreated cells (P < 0.05). Conclusion: Over-expression of Tiam-1 contributes to the invasive phenotype of gastric cancer cells. Inhibition of Tiam 1 expression could impair the invasive capacity of gastric cancer cells through modulating reconstruction of the cytoskeleton and regulating expression of MMP 2.

• Reproductive and Developmental Biology
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• 제37권3호
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• pp.103-108
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• 2013

An uterus is female reproductive tract organ that affected estrus cycle. During a various changes occur at uterus in estrus cycle, one of them is body fluids secretion be called uterine fluid. Therefore, the objective of this study was to investigate the changes of protein patterns using two-dimensional gel electrophoresis in uterus fluids during the follicular and luteal phases in estrus cycle of pigs. In changes of protein spots were confirmed during the follicular and luteal phases. The 136 spots were expressed in follicular phase, the 57 spots of them showed reproducibility. On the other hand, the 140 spots were expressed in luteal phase, the 73 spots of them showed reproducibility. Also, spots expressed in follicular phase were number 69 and 94 spots and spots expressed in luteal phase only were number 156, 157, 184~187, 190 and 191 spots. The spots which of higher expression levels in the luteal phase than in follicular phase were number 76 and 79 spots. In conclusion, the spots expressed in follicular and luteal phases were confirmed with difference levels and these differences are function of RNA resolving, protein synthesis and cytoskeletal architecture.