• Title/Summary/Keyword: Microfilament

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Microtubule and Microfilament Dynamics in Porcine Oocytes during Meiotic Maturation, Fertilization and Parthenogenesis (돼지 난자의 성숙, 수정 및 단위발생시 Microtubule과 Microfilament의 움직임)

  • 김남형;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.19 no.3
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    • pp.205-216
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    • 1995
  • Microtubules와 micrfilaments는 포유동물 난자이 주요한 세포 구조물들로, 이들은 난자의 성숙, 수정 및 배발달시 핵질의 이동과 세포질 분열에 직접 관여하는 것으로 알려져 왔다. 난자내 세포구조물의 정확한 움직임은 정상적인 배 발달을 위해 필수적이다. Microtubules는 $\alpha$, $\beta$- bubulin이 서로 연결되어 이루어져 있으며, 수정시 웅성.자성전핵 움직임과 세포분열시, 유사 및 감수분열시 그 역할을 한다. 생쥐를 제외한 대부분의 동물에서 microbubules의 역할은 수정시 정자가 centrosome을 난자내로 이전하여 sperm aster를 형성함으로써 시작된다고 보고되고 있다. 따라서 정자의 도움없이 배발달이 일어나는 단위발생시 microbubules의 형성은 연구들 사이에 흥미로운 연구대상이 되고 있다. 한편 microfilaments는 세포분열시 세포질을 분할하는 기계적인 역할을 하는 것으로 알려져 있으며, 최근 생쥐 난자에서는 정자의 난자내 융합과 웅성 및 자성 전핵의 이동에 관여한다고 보고되고 있다. 포유동물 난자의 체외성숙, 체외수정을 유도할 때 여러 가지 비정상적인 핵움직임과 세포분열이 관찰되어지고, 낮은 배발달율이 보고되고 있는데, 최근 연구자들은 세포구조물, 즉 microtubules와 microfilaments의 비정상적인 역할에서 기인한다고 보고 있다. 따라서 포유동물 난자의 성숙.수정 및 단위발생시 세포구조물의 움직임과 역할 및 상호관계에 대한 정확한 이해는 체외수정율 및 배발달 향상에 중요한 기초자료로 이용되리라고 본다.

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Morphological Characteristics of Sperm in the Korean Striped Field Mouse, Apodemus agrarius coreae: Possible Role of Sperm Neck in the Movement of Sperm Head

  • Lee, Jeong-Hun;Son, Seong-Won
    • Animal cells and systems
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    • v.1 no.2
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    • pp.371-379
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    • 1997
  • To investigate the movement of sperm head and the role of sperm neck in forward sperm motility in the Korean striped field mouse, Apodemus agrarius coreae, the morphological characteristics of the cauda epididymal spermatozoa were examined by light microscopy and scanning and transmission electron microscopy. Spermatozoa of A. agrarius coreae were characterized by the conspicuous shape of the acrosome and the long tail compared with those of other rodents. Total length of the sperm was $133\mu{m}$. The sperm head had a curved falciform shape. The head was 8.0${\mu}$m in length, and about 4.0 ${\mu}$m in width. The shape of acrosome had an openerlike form. The sperm tail (125 ${\mu}$m) consisted of four major segments: neck (0.5 ${\mu}$m), middle piece (29.5 ${\mu}$m), and principal piece plus the end piece (95 ${\mu}$m). The outer dense fibers were arranged in a horseshoe fashion, and No. 1, 5, 6, and 9 of the outer dense fibers were larger than the others. The mitochondrial bundles of middle piece were composed of a pair of arms, which surrounded the axone of the middle piece by the 45 0 angled helical structure. The total number of mitochondrial gyres was 188. In particular, the microfilament structures existed in plasma membrane of the sperm, which was adjacent to the acrosomal region on the nuclear membrane. The segmented columns were surrounded by microfilament structures, and the microfilament bundles were adjacent to the outer membrane of the first mitochondria of middle piece. This study presents for the first time the existence of microfilament structures within the plasma membrane of sperm which is located from the adjacent acrosome region to the connecting piece in sperm neck of Korean striped field mouse, Apodemus agrarius coreae. The present result suggests that the constriction and extension of microfilament in sperm neck as well as the wave-movement of sperm tail may play a role in the movement of sperm head.

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Distribution patterns of cytoskelectal proteins in cardiac endothelial cells : Investigation using monoclonal antibodies (심장내피세포 골격단백의 배열 양상 : monoclonal antibodies를 이용한 조사)

  • Kim, Han-Chul;Song, In-Hwan;Lee, Yung-Chang
    • Journal of Yeungnam Medical Science
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    • v.7 no.2
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    • pp.27-37
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    • 1990
  • To investigate the changing patterns of microfilament and microtubule arrangement and influence of myocardial cells and colchicine to microfilament and microtubule formation in cardiac endothelial cells the authors carried out indirect immunofluorescence stain for actin and tubulin with supernatant monoclonal antibodies. Secondary antibodies were IgG FITC conjugate. The results were summerized as follows. Fiberform reactions were stronger in the cells with many processes and spread cytoplasm and they became weaker after the endothelial cells formed monolayer. In the endothelial cells cocultured with myocardial cells the fiberform of the microtubule became less visible compared to control group but fiberform of the microtubule maintained strong intensity as endothelial cells formed monolayer. In the group treated with colchicine, there were no visible differences in microfilaments compared to control group but fiberform of microtubule revealed weaker intensity after colchicine treatment. The intensity of microtubule fiberform returned to control level after 2 days.

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The effect of microfilament inhibitor on the Cryptosporidium infection in vitro

  • Yu, Jae-Ran;Choi, Saung-Don
    • Parasites, Hosts and Diseases
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    • v.38 no.4
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    • pp.257-261
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    • 2000
  • This study was focused on the effects of microfilament inhibitor, Cytochalasin D (CD) on the invasiveness of sporozoites of Cryptosporidiun spp. into the host cells. MDCK and AGS cell lines were used as host cells for C. parvum and C. muris, respectively. When MDCK cells were pretreated with CD for 1 hr before inoculation of the sporozoites, C. parvum infection was significantly inhibited when compared to the control cells. These inhibitory effects of CD on the rate of infection were dose-dependent. In addition, C. muris infection was hampered when AGS cell lines were pretreated with CD. However, the capability of invasiveness of the sporozoites into the host cells was not greatly influenced by the pretreatment of sporozoites with CD before infection. These results suggest that microfilaments of host cells, rather than parasites, play an important role for the invasion of Cryptosporidium spp.

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Cortical Granule Distribution During In Vitro Maturation and Fertilization of Porcine Oocytes (돼지난자의 체외성숙 및 수정시 일어나는 표층과립막세포의 분포변화에 관한 연구)

  • 송상진;권중균;도정태;김남형;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.343-351
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    • 1996
  • The objectives of this study are to determine cortical granule distribution during in vitro maturation, parthenogenetic activation and in vitro fertilization of oocytes, and to investigate effects of microfilament inhibitor on the cortical granule distribution during in vitro maturation and fertilization of oocytes in the pig, The corti-cal granule distribution were imaged with fluor-escent labeled lectin under laser scanning confocal microscope or detected by transmission electron microscope. At germinal vesicle stage, cortical granule organelles were located around the cell cortex and were present as a relatively thick area on the oolema. Microfilaments were also observed in a thick uniform area around the cell cortex. Following germinal vesicle break down,microfilaments concentrated to the condensed chromatin and cortical granules were observed in the cortex. Treatment with cytochalasin B inhibited microfilament polymerization and prevented movement of cortical granules to the cortex. Cortical granule exudate following sperm penetration was evenly distributed in the entire perivitelline space. Therefore, these results suggested that the microfilament assembly is involved in the distribution, movement and exocytosis of cortical granules during maturation and fertilization of porcine oocytes. (Key words cortical granule, porcine, maturation, fertilization).

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The effects of the surrounding viscoelastic media on the buckling behavior of single microfilament within the cell: A mechanical model

  • Khadimallah, Mohamed A.;Safeer, Muhammad;Taj, Muhammad;Ayed, Hamdi;Hussain, Muzamal;Bouzgarrou, Souhail Mohamed;Mahmoud, S.R.;Ahmad, Manzoor;Tounsi, Abdelouahed
    • Advances in concrete construction
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    • v.10 no.2
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    • pp.141-149
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    • 2020
  • In the present study, a mechanical model is applied to account the effects of the surrounding viscoelastic media on the buckling behavior of single microfilament within the cell. The model immeasurably associates filament's bending rigidity, neighboring system elasticity, and cytosol viscosity with buckling wavelengths, buckling growth rates and buckling amplitudes of the filament. Cytoskeleton components in living cell bear large compressive force and are responsible in maintaining the cell shape. Actually these filaments are surrounded by viscoelastic media consisting of other filaments network and viscous cytosole within the cell. This surrounding, viscoelastic media affects the buckling behavior of these filaments when external force is applied on these filaments. The obtained results, indicate that the coupling of viscoelastic media with the viscous cytosol greatly affect the buckling behavior of microfilament. The buckling forces increased with the increase in the intensity of surrounding viscoelastic media.

Effects of Alkylating Agent on the Sertoli Cell of the Seminiferous Tubule in the Mouse (Alkylating agent가 생쥐 정소의 Sertoli Cell에 미치는 영향)

  • Jung, Hae-Man;Cho, Kwang-Phil;Kim, Jeong-Sang
    • Applied Microscopy
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    • v.26 no.3
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    • pp.293-303
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    • 1996
  • This paper aims to probe that the effect of high dose of cyclophosphamide to the Sertoli cells of the mouse was examined by transmission electron microscope. In the normal group, Sertoli cells contact each other around the basal aspect of the seminiferous tubule, forming numerous row of tight junction, blood-testis barrier. Sertoli cells contain smooth endoplasmic reticulum, well developed Golgi comples, a number of round mitochondria and microfilament. The cytoplasmic necrosis are observed from the 1-time treated group. In the 2-times treated group, smooth endoplasmic reticulum are more developed than normal group, but cisternae are partially dilated. In the 3-times treated group, the smooth endoplasmic reticulum are not developed. In the 2-times treated group, the inner membrane of the mitochondria are partially disrupted, and cristae are all disrupted in the 3-times treated group. The microfilaments are not observed in the all treated groups. According to the results above, it seems that smooth endoplasmic reticulum, mitochondria, and microfilament are disrupted by toxic effects of the cyclosphamide to the Sertoli cells of the mouse.

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The Transfection of Caldesmon DNA into Primary Cultured Rat Aortic Vascular Smooth Muscle

  • Choi, Woong;Ahn, Hee-Yul
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.6
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    • pp.597-603
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    • 1999
  • Caldesmon (CaD), one of microfilament-associated proteins, plays a key role in microfilament assembly in mitosis. We have investigated the effects of overexpression of the high molecular weight isoform of CaD (h-CaD) on the physiology of vascular smooth muscle cells (VSMCs). Rat aortic VSMCs were stably transfected with plasmids carrying a full length human h-CaD cDNA under control of cytomegalovirus promoter. The majority of the overexpressed h-CaD appears to be localized predominantly on cytoskeleton structures as determined by detergent lysis. The overexpression of h-CaD, however, does not decrease the level of endogenous low molecular weight isoform of CaD. h-CaD overexpressing VSMCs (h-CaD/VSMCs) show a decreased growth rate than that of vector-only transfected cells when determined by $[^3H]thymidine$ uptake and cell counting after fetal bovine serum (FBS) stimulation. h-CaD/VSMCs were smaller than vector-transfected cells by 18% in cell diameter. These data suggest that overexpression of h-CaD can inhibit the poliferation and the cell volume of VSMCs stimulated by growth factors and that the gene therapy with h-CaD may be helpful to prevent the conditions associated with hypertrophy and/or hyperplasia of VSMCs after arterial injuries.

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The Cytoskeletal and Chromosomal Constitution in Vitrified Mouse Oocyte (초자화동결된 생쥐 미수정란의 Cytoskeleton 및 염색체 변화)

  • Yi, B.K.;Kim, E.Y.;Nam, H.K.;Lee, K.S.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.3
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    • pp.287-292
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    • 1998
  • The objective of this study was to confirm whether the vitrification method using EFS35 has detrimental effect for cytoskeleton and chromosome constitution of the mouse oocytes by indirect immunocytochemistry and chromosome analysis. Mouse oocytes were vitrified using EFS35 which consisted of 35% ethylene glycol, 18% ficoll, 0.3 M sucrose and 10% FBS in M2 medium. The results obtained in this experiment were summarized as follows: When the survival rates after being exposed or vitrified in EFS35 were examined, there were not different between two groups (97.7 and 89.3%). Also, when the microtubule morphology and microfilament distribution in vitrified oocytes were examined, normal percentage of two cytoskeleton in vitrified group (95.5 and 100%) was not different from that in control (97.5 and 100%) and exposed group (92.3 and 100%). In addition, the rate of oocytes containing a normal chromosome number in vitrified group (73.5%) after IVF was not different from that in control (79.5%) and exposed group (78.7%). These results indicated that the cytoskeletal morphology and chromosome constitution of mouse oocytes were not affected by cryoprotectant (EFS35) or freezing apd that vitrification methods using EFS35 was suitable for cryopreservation of mouse oocytes.

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