• Journal of Life Science
• /
• v.31 no.3
• /
• pp.280-286
• /
• 2021

Oryzalin is a herbicide that disrupts the arrangement of microtubules by binding to tubulin, thereby blocking the anisotropic growth of plant cells. Microtubules and microfilaments are cytoskeleton components that have been implicated in plant growth through their influence on the formation of cell walls. Microtubules also play roles in the sedimentation of amyloplasts in the root tip columella cells; this sedimentation is related to gravity sensing and results in downward root growth in the soil for absorption of water and minerals. However, the orientation of microtubules changes depending on the level of ethylene in plant cells. A recent study reported that oryzalin stimulated ethylene production via 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase and caused a concentration-dependent inhibition of root growth and gravitropic responses. The aim of the present study was to investigate the possibility that oryzalin-induced inhibition might be recovered by the application of inhibitors of ethylene production, such as 10-4 M cobalt ions and 10-8 M aminoethoxyvinylglycine (AVG). The inhibition of root growth and gravitropic response was overcome by 10-20% by an 8 hr treatment with cobalt ions or AVG. These results suggest that ethylene levels could regulate root growth and gravitropic responses in Arabidopsis.

• Journal of Life Science
• /
• v.31 no.2
• /
• pp.109-114
• /
• 2021

Oryzalin is a dinitroaniline herbicide that has been known to disrupt microtubules. Microtubules and microfilaments are components of cytoskeletons that are implicated in plant cell growth, which requires the synthesis of cellulose when cell walls elongate. In addition, microtubules are also involved in the sedimentation of statoliths, which regulate the perception of gravity in the columella cells of root tips. In this study, we investigated the effect of oryzalin on the growth and gravitropic response of Arabidopsis roots. The role of ethylene in oryzalin's effect was also examined using these roots. Treatment of oryzalin at a concentration of 10-4 M completely inhibited the roots' growth and gravitropic response. At a concentration of 10-6 M oryzalin, root growth was inhibited by 47% at 8 hr when compared to control. Gravitropic response was inhibited by about 38% compared to control in roots treated with 10-6 M oryzalin for 4 hr. To understand the role of oryzalin in the regulation of root growth and gravitropic response, we measured ethylene production in root segments treated with oryzalin. It was found that the addition of oryzalin stimulated ethylene production through the activation of ACC oxidase and ACC synthase genes, which are key components in the synthesis of ethylene. From these findings, it can be inferred that oryzalin inhibits the growth and gravitropic response of Arabidopsis roots by stimulating ethylene production. The increased ethylene alters the arrangement of the microtubules, which eventually interferes with the growth of the cell wall.

• Applied Microscopy
• /
• v.28 no.1
• /
• pp.39-48
• /
• 1998

The spermatozoa of bagrid catfish, Pseudobagrus fulvidraco are approximately $76{\mu}m$ in length, and a relatively simple and elongated cell composed of a spherical head, a short middle piece and a tail. The ultrastructure of spermatozoa of P. fulvidraco is characterized by the following features. The acrosome is absent as in most teleost. The round nucleus measuring about $1.67{\mu}m$ in length and diameter is depressed with a deep nuclear fossa. The nuclear fossa, the length of which is about three-fifths of the nuclear diameter, contains the proximal and distal contrioles. The two centrioles are oriented approximately $160^{\circ}$ to each other. The filamentous materials give rise to satellite appendages arranged tangentially from the triplets of the distal centriole and the doublets of the anterior end of the axoneme toward the nuclear envelope. The mitochondria are not fused and their number is 20 or more. They are arranged in two or three layers and two rings within the cytoplasmic collar and surround the axoneme. They are separated from the axoneme by the cytoplasmic canal. The axoneme is of the 9+2 microtubular pattern and has inner but no outer dynein arms. The two lateral fins are in the same plane with the two central microtubules, the doublets 3 and 8, which are ultrastructural characteristics of the sperm tail unlike other siluroids lacking the lateral fins.

• Journal of Life Science
• /
• v.27 no.6
• /
• pp.673-679
• /
• 2017

Microtubules are long rods in the cytoplasm of cells that plays a role in cell motility and intracellular transport. Microtubule-based transport by motor proteins is essential in intracellular transport. Kinesin 1 is a molecular motor protein that mediates the intracellular transport of various membranous vesicles, mRNAs, and proteins along microtubules. It is comprised of two heavy chains (KHCs, also called KIF5s) and two light chains (KLCs). KIF5s bear a motor domain in their amino (N)-terminal regions and interact with various cargoes through the cargo-binding domain in their carboxyl (C)-terminal regions. To identify proteins interacting with KIF5B, yeast two-hybrid screening was performed, and a specific interaction with the cytoplasmic linker protein 170 (CLIP-170), a plus end microtubule-binding protein, was found. The coiled-coil domain of CLIP-170 is essential for interactions with KIF5B in the yeast two-hybrid assay. CLIP-170 bound to the cargo-binding domain of KIF5B. Also, other KIF5s, KIF5A and KIF5C, interacted with CLIP-170 in the yeast two-hybrid assay. In addition, glutathione S-transferase (GST) pull-downs showed that KIF5s specifically interacted with CLIP-170. An antibody to KIF5B specifically co-immunoprecipitated CLIP-170 associated with KIF5B from mouse brain extracts. These results suggest that kinesin 1 motor protein may transport CLIP-170 in cells.

• Journal of Life Science
• /
• v.25 no.11
• /
• pp.1223-1229
• /
• 2015

Oryzalin is a dinitroaniline herbicide, which disrupts the arrangement of microtubules. Microtubules and microfilaments are cytoskeletal components that are thought to play a role in the sedimentation of statoliths and the formation of cell walls. Statoliths regulate the perception of gravity by columella cells in the root tip. To determine the effect of oryzalin on the gravitropic response, ethylene production in primary roots of maize was investigated. Treatment with 10^-4 M oryzalin to the root tip inhibited the growth and gravitropic response of the roots. However, the treatment had no effect on the elongation zone of the roots. An application of 10^-4 M oryzalin for 15 hr to the root tip caused root tip swelling. The application of 1-aminocycopropane-1-carboxylic acid (ACC), a precursor of ethylene, to the root tip also inhibited the gravitropic response. To understand the role of oryzalin in the regulation of the growth and gravitropic response of roots, ethylene production in the primary roots of maize was measured following treatment with oryzalin. Oryzalin stimulated ethylene production via the activation of ACC oxidase (ACO) and ACC synthase (ACS), and it increased the expression of ACO and ACS genes. Indole-3-acetic acid (IAA) played a key role in the asymmetric elongation rates observed during gravitropism. The results suggest that oryzalin alters the gravitropic response of maize roots through modification of the arrangement of microtubules. This might reduce the distribution of IAA in the upper and lower sides of the elongation zone and increase ethylene production, thereby inhibiting growth and gravitropic responses.

• Journal of Life Science
• /
• v.32 no.2
• /
• pp.161-166
• /
• 2022

Mitosis is a process in which a replicated genome is distributed to two daughter cells, and it is necessary for cell survival and organismal development. During mitosis, the spindle assembly checkpoint (SAC) ensures faithful chromosome segregation by monitoring the kinetochore attachment to the mitotic spindle. Although the SAC mechanism has been extensively studied over the last 30 years, the mechanism by which a single unattached kinetochore activates the SAC remains unclear. The key components of the SAC are Mad1, Mad2, Mad3 (BubR1 in higher eukaryotes), Bub1, Bub3, and Cdc20, which are all required for SAC activation. An essential step for SAC activation is the formation of the Mad2 - Cdc20 complex in the unattached kinetochore, which is kinetically disfavored. Although the mechanism by which Mad2 and Cdc20 are recruited to unattached kinetochores is well-known, it is not clear how they form a complex. Recently, a key mechanism for the formation of the Mad2 - Cdc20 complex has been identified, which is catalyzed by an unattached kinetochore. This supports the evidence that a single unattached kinetochore can activate the SAC signaling. Herein, we discuss the known key mechanism for SAC activation, review the recent studies on SAC, and conclude how their discoveries improved the understanding of mitosis.

• Applied Microscopy
• /
• v.39 no.2
• /
• pp.185-189
• /
• 2009

Studies about the structure of Drosophila melanogaster retinal cell using electron microscopy have been carried out in details since 1960s. However, these results can have limitations in functional research because of two-dimensional structure. In this study, the adult retina of Drosophila melanogaster was investigated by employing high pressure freezing method, serial sections, high voltage electron microscopy, and 3-dimensional reconstruction method. From there results, mitochondria, microtubules, and nuclei were reconstructed as 3-dimensional structure using IMOD program. The 3D structure of these organelles showed that mitochondira mainly located in distal region near lens, and microtubule mainly located in distal and basal region. The 3D reconstruction of these organelles can be used for a critical evaluation in the dynamic change of cellular organelles caused by functional abnormality like retinal degeneration.

• Journal of Life Science
• /
• v.21 no.8
• /
• pp.1076-1082
• /
• 2011

Microtubule-based kinesin motor proteins are used for long-range vesicular transport. KIF5s (KIF5A, KIF5B and KIF5C) mediate the transport of various membranous vesicles along microtubules, but the mechanism behind how they recognize and bind to a specific cargo has not yet been completely elucidated. To identify the interaction protein for KIF5B, yeast two-hybrid screening was performed and a specific interaction with the unconventional myosin Myo9b, an actin-based vesicle transport motor, was found. The GTPase-activating protein (GAP) domain of Myo9s was essential for interaction with KIF5B in the yeast two-hybrid assay. Myo9b bound to the carboxyl-terminal region of KIF5B and to other KIF5 members. In addition, glutathione S-transferase (GST) pull-downs showed that Myo9s specifically interact to the complete Kinesin-I complex. An antibody to KIF5B specifically co-immunoprecipitated KIF5B associated with Myo9s from mouse brain extracts. These results suggest that kinesin-I motor protein interacts directly with actin-based motor proteins in the cell.

• Applied Microscopy
• /
• v.1 no.1
• /
• pp.35-42
• /
• 1969

가토간장에서 채취(採取)한 간(肝)디스토마 성충(成蟲)의 정소(精巢)와 수정낭(受精囊)을 1.25% glutaraldehyde와 1% 사산화(四酸化) 오스뮴산(酸)으로 냉실(冷室)에서 이중고정(二重固定)하였다. 고정(固定)된 시료(試料)는 양식(樣式)에 따라 탈수(脫水)한 후(後) Epon-812로 포매(包埋)하여 MT-2형(型) Porter Blum microtome으로서 초박절편(超薄切片)을 만들어 수사화연(水酸化鉛)과 초산(醋酸)우라닐로서 이중염색(二重染色)한후 Hitachi HS-7형(型)과 HU-11E형(型)의 전자현미경(電子顯微鏡)으로 관찰(觀察)하였다. 관찰결과 간(肝)디스토마의 정세포(精細胞)는 타원형으로 난형(卵形)의 인(仁)을 포함(包含)한 큰 핵(核)을 갖고 있으며 핵(核)을 둘러싼 비교적소량(比較的小量)의 세포질(細胞質)에는 낭형(囊形)의 조면소포체(粗面小胞體)가 희소(稀少)하게 있으며 유리(遊離)리보좀 및 즐(櫛)을 가진 미토콘드리아, 중심체(中心體), 층판상(層板狀)의 골지체가 있다. 정자(精子)는 긴 원주형(圓柱形)의 핵(核)과 선단(先端)에 첨체(尖體)를 포함(包含)한 두부(頭部)와 중종편(中終片), 미부(尾部)의 말단부(末端部)로 갈수록 점차 가늘어져서 결국(結局) 편모(鞭毛)로 끝난다. 두부(頭部)의 선단부(先端部) 가까이 핵환(核環)이 있으므로 이를 기시점(起始點)으로 직경(直徑) $250{\AA}$ 정도의 $8{\sim}10$개(個)의 미세소관(微細小管)이 축사(軸絲)의 배면(背面) 원형질막(原形質膜) 직내면(直內面)의 외형질(外形質)에 평행(平行)하게 중종편(中終片)까지 신장(伸長)되어 있다. 미토콘드리아는 융합(融合)되어 두부(頭部)의 후반부(後半部) 핵(核)의 복측(腹側)에 평행(平行)하게 축사(軸絲)를 감싸는 원추형(圓錐形)으로 종단면(縱斷面)에서 반점상(班點狀)의 횡문(橫紋)이 관찰(觀察)되었다. 중심체(中心體)에서 기원(起原)된 축사(軸絲)는 중심부(中心部)에 중심섬유(中心纖維)와 중심초 및 이중(二重)의 미세소관(微細小管)이 9개(個) 둘러싸고 있는 구조(構造)를 하고 있다.

• Korean Journal of Microbiology
• /
• v.36 no.4
• /
• pp.306-311
• /
• 2000

Infection of Vero cells with herpes simplex virus type-1 (HSV-1) resulted in a series of changes in intra-cellular free calcium concentration $([Ca^{2+}]_i)$. A significant and maximal decrease $[Ca^{2+}]_i$ was observed at 4 hours postinfection (hr p.i.) in HSV-1-infected in Vero cells. Inactivation of HSV-1 with UV irradiation and heat treatment abolished HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells. And the degree of the decrease in $[Ca^{2+}]_i$ was dependent on the amount of input virus. Taxol, which stabilizes the polymerization of microtubule blocked HSV-1-induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i., suggesting that microtubule may mediate the transport of HSV-1 nucleocapsid to the nucleus of infected cell. Treatment of HSV-1-infected Vero cells with metabolic inhibitors such as cycloheximide, cordycepin, or acyclovir partially reversed the decrease in $[Ca^{2+}]_i$ at 4 hr p.i.. Thus, it is suggested that HSV-1 induced decrease in $[Ca^{2+}]_i$ at 4 hr p.i. in Vero cells may play an important role in the multiplication of HSV-1.