• Title/Summary/Keyword: ${\gamma}$-tubulin

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Inhibition of Gastric Cancer Cell Cycle Progression by ${\gamma}$ -Tubulin Antisense Oligonucleotides

  • Hwang, Sun-Hee;Kim, Myung-Won;Park, Sang-Kyu;Noh, Jung-Woo;Han, In-Seob
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.876-879
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    • 2001
  • ${\gamma}$ -Tubulin is an essential component involved in microtubule nucleation. The present work examined whether the fast proliferation of cancer cells can be retarded by the depletion of ${\gamma}$ -tubulin expression. Two different gastric cancer cell lines and one control cell line were treated with antisence oligonucleotides complementary to the messenger RNA of ${\gamma}$ -tubulin. The$[^3H]$ -thymidine incorporation in the two gastric cancer cell lines, SNU-1 and SNU-216, was dramatically reducd by treatment with the ${\gamma}$ -tubulin antisense oligonucleotides in a dosage-dependent manner. In contrast, the control cell line, NIH/3T3, showed no significant effect from the antisense oligonucleotides even at a high concentration. The ablation of ${\gamma}$ -tubulin expression in the tumor cells resulted in an altered DNA synthesis during mitosis and it decreased the cell progression. Accordingly, the use of antisense oligonucleotides may be an effective way of inhibiting the proliferation of human gastric cancers.

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Gamma (${\gamma}$) Tubulin Reorganization during Bovine Fertilization and Parthenogenesis

  • Yung, Youn-Ji;Shin, Mi-Ra;Lee, Hwa-Young;Park, Sang-Hyun;Cui, Xiang-Sun;Kim, Nam-Hyung
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.49-49
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    • 2002
  • γ - tubulin is an essential, invariant constitutive centrosomal protein, which plays key roles in microtubule patterning and defining the microtubule intrinsic polarity. Although γ-tubulin was also present in cattle oocytes and zygotes, no details have been provided on its recruitment and localization to date. In this study, we determined γ-tubulin distribution chronologically in conjunction with microtubule dynamics during fertilization and parthenogenesis, with a view to understanding the molecular basis of zygotic centrosome reconstitution in cattle. (omitted)

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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.

Fate of Donor Centrosome and Microtubule Dynamics of Porcine Somatic Cell Nuclear Transfer Embryos

  • Kwon, Dae-Jin;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.34 no.2
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    • pp.73-79
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    • 2010
  • We investigated the microtubule dynamics, including the inheritance of donor centrosomes and the mitotic spindle assembly occurring during the first mitosis of somatic cell nuclear transfer (SCNT) embryos in pigs. SCNT embryos were fixed 15 min and 1 h after fusion in order to assess the inheritance pattern of the donor centrosome. The distribution and dynamic of the centrosome and microtubule during the first mitotic phase of SCNT embryos were also evaluated. The frequency of embryos evidencing $\gamma$-tubulin spots (centrosome) was 93.2% in the SCNT embryos 15 min after fusion. In the majority of the SCNT embryos (61.5%), however, no centrosome was observed 1 h after fusion. The frequency of the embryos with no or abnormal mitotic spindles 20 h after fusion was 19.6%. The $\gamma$-tubulin spots were detected near the nuclei of somatic cells regardless of cell cycle phase, whereas $\gamma$-tubulin spots in the SCNT embryos were observed only during the inter-anaphase transition. These results showed that the donor centrosome is inherited into the SCNT embryos, but failed to assemble the normal mitotic spindles during first mitotic phase in some SCNT embryos.

Gamma ($\GAMMA$) Tubulin and Microtubule Assembly in Porcine Nuclear Transfered Embryos and Parthenotes

  • Hong, Jun-Soon;Park, Sang-Hyun;Shun, Cui-Xiang;Kim, Nam-Hyung
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.51-51
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    • 2002
  • Despite of importance of integrated events of nucleus and microtubule remodeling in nuclear transferred embryos with somatic cells, little information is available on this subject. In this study we compared chromatin, r-tubulin and microtubule organization in porcine oocytes following somatic cell nuclear transfer and parthenogenetically activation in order to clarify nuclear remodeling process and to demonstrate centrosome inheritance during nuclear transfer. (omitted)

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An Immunohistochemical Study of Pheasant Testis in Active, Inactive and Damaged States (번식기, 비번식기 및 손상상태에 따른 한국꿩 고환의 면역조기화학적 연구)

  • ;Karl-heinz Wrobel
    • Korean Journal of Poultry Science
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    • v.24 no.3
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    • pp.107-116
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    • 1997
  • In order to achieve optimal reproductive performance, reliable morphological and physiological basic data on the reproductive organs are desirable. Adult male Korean ring-necked pheasant in inactive(mid of January) and active state (end of April) were used in this study. In addition, five active state pheasants were received a single dose of 60Co-ray 500 rads each to damage the testes. The objective of this study was to investigate the distribution pattern of protein gene product (PGP) 9.5 and ${\alpha}$-tubulin in the pheasant testes of the active, inactive and ${\gamma}$-ray irradiated active states. The results obtained were summarized as follows 1. The seminiferous tubules collected in inactive states( mid of Jan) showed narrow lumen, and the spermatogonia and the Sertoli cell were well preserved. The PGP 9.5 immunoreactivity of these tubules showed a positive reaction in paranucleus area of the spermatogonia, and a positive reaction in a small number of the Leydig cells in the interstitium of the seminiferous tubules. 2. The seminiferous tubules were dilated in active state(end of April) as compared with the inactive state. The PGP 9.5 reactivity in these tubules showed a positive reaction in many Leydig cells in the interstitium of the seminiferous tubules, and the testes of ${\gamma}$-ray irradiated group showed partially weak reaction in the interstitium of the seminiferous tubules. 3. The ${\alpha}$-tubulin reactivity in the seminiferous tubules of the inactive testes was strongly positive in the cytoplasmic process of the Sertoli cell from the basal stem region to the apical ex-tension. From the broad part of the stem region to the luminal space, the active testes showed a strong positive reaction. The ${\gamma}$-ray irradiated groups showed diminished reaction in the basal region.

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A Tubulin Inhibitor, N-(5-Benzyl-1,3-thiazol-2-yl)-3-(furan-2-yl)prop-2-enamide, Induces Anti-inflammatory Innate Immune Responses to Attenuate LPS-mediated Septic Shock

  • Park, Hyun Jung;Lee, Sung Won;Park, Hwangseo;Park, Se-Ho;Hong, Seokmann
    • Bulletin of the Korean Chemical Society
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    • v.35 no.11
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    • pp.3307-3312
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    • 2014
  • The anti-inflammatory effect of a tubulin inhibitor, N-(5-benzyl-1,3-thiazol-2-yl)-3-(furan-2-yl)prop-2-enamide (1), on innate immune responses remains unclear. Thus, we investigated the effect of 1 on the immune responses mediated by lipopolysaccharide (LPS). The in vitro addition of 1 to dendritic cells and macrophages dose-dependently reduced tumor necrosis factor alpha production elicited by LPS stimulation. Additionally, the stimulation of natural killer (NK) and natural killer T (NKT) cells with 1 resulted in the decrease of interferon gamma ($IFN{\gamma}$) induced by LPS treatment. Moreover, 1 substantially reduced interleukin 12 in dendritic cells (DC) as well as $IFN{\gamma}$ in NKDCs induced by LPS in vitro. Furthermore, the in vivo administration of 1 ameliorated LPS/D-galactosamine-induced endotoxic lethality in mice. Taken together, our results demonstrate for the first time that 1 possesses anti-inflammatory properties, most notably by modulating LPS-induced innate immune responses. Therefore, 1 might have therapeutic potential for the treatment of inflammation-mediated diseases such as sepsis.

Microtubule Assemblies and Methylation Patterns of Porcine IVF and Parthenogenetic Embryos (돼지 체외수정란 및 단위발생란의 미세관 형성 및 메틸화 양상)

  • Park, Joo-Hee;Kim, Ho-Jeong;Lee, Beom-Ki;Kwon, Dae-Jin;Hwang, In-Sun;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.7-11
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    • 2009
  • This study was performed to confirm the microtubule assemblies and methylation patterns of porcine IVF and parthenogenetic embryos. Cumulus-oocyte complexes were collected and matured in vitro for 42 hr. Oocytes were fertilized by prepared fresh sperm or activated parthenogenetically by exposure to electric stimulation and 6-dimethylaminopurine. Porcine IVF and parthenogenetic embryos were cultured in vitro for 6 days. Embryos were stained by immunofluorescence staining method to observe the dynamic of nucleus and microtubules in the first mitotic phase and the methylation patterns in different developmental stages. After then, samples were confirmed and analyzed through a laser-scanning confocal microscope. IVF embryos had a centrosome originated from sperms, which was shown a $\gamma$-tubulin spot. However, $\gamma$-tubulin spot was not observed in parthenogenetic embryos. A lower methylation level was observed in IVF embryos compared to parthenogenetic ones at the morula and blastocyst stages. In conclusion, it is considered that microtubule assemblies and genetic regulation mechanism differ between parthenogenetic and IVF embryos.