ジャーナル: Nat Cell Biol / 年: 2015 タイトル: Quantitative analysis of cytoskeletal reorganization during epithelial tissue sealing by large-volume electron tomography. 著者: Mikhail Eltsov / Nadia Dubé / Zhou Yu / Laurynas Pasakarnis / Uta Haselmann-Weiss / Damian Brunner / Achilleas S Frangakis / 要旨: The closure of epidermal openings is an essential biological process that causes major developmental problems such as spina bifida in humans if it goes awry. At present, the mechanism of closure ...The closure of epidermal openings is an essential biological process that causes major developmental problems such as spina bifida in humans if it goes awry. At present, the mechanism of closure remains elusive. Therefore, we reconstructed a model closure event, dorsal closure in fly embryos, by large-volume correlative electron tomography. We present a comprehensive, quantitative analysis of the cytoskeletal reorganization, enabling separated epidermal cells to seal the epithelium. After establishing contact through actin-driven exploratory filopodia, cells use a single lamella to generate 'roof tile'-like overlaps. These shorten to produce the force, 'zipping' the tissue closed. The shortening overlaps lack detectable actin filament ensembles but are crowded with microtubules. Cortical accumulation of shrinking microtubule ends suggests a force generation mechanism in which cortical motors pull on microtubule ends as for mitotic spindle positioning. In addition, microtubules orient filopodia and lamellae before zipping. Our 4D electron microscopy picture describes an entire developmental process and provides fundamental insight into epidermal closure.
ダウンロード / ファイル: emd_2610.map.gz / 形式: CCP4 / 大きさ: 1 GB / タイプ: IMAGE STORED AS SIGNED BYTE
注釈
Tomographic reconstruction of Drosophila zipping
ボクセルのサイズ
X: 59 Å / Y: 59 Å / Z: 81 Å
密度
最小 - 最大
-128.0 - 127.0
平均 (標準偏差)
1.30015326 (±30.184339520000002)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
957
-52
14
サイズ
999
892
1254
Spacing
999
892
1254
セル
A: 52628.0 Å / B: 58941.0 Å / C: 101574.0 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
envelope stored as signed bytes (from -128 lowest to 127 highest)
Å/pix. X/Y/Z
59
59
81
M x/y/z
892
999
1254
origin x/y/z
0.000
0.000
0.000
length x/y/z
52628.000
58941.000
101574.000
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
-52
957
14
NC/NR/NS
892
999
1254
D min/max/mean
-128.000
127.000
1.300
-
添付データ
-
試料の構成要素
-
全体 : Tomographic reconstruction of an entire zipping in Drosophila embryo
全体
名称: Tomographic reconstruction of an entire zipping in Drosophila embryo
要素
試料: Tomographic reconstruction of an entire zipping in Drosophila embryo
細胞器官・細胞要素: zipping in Drosphila embryogenesis
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超分子 #1000: Tomographic reconstruction of an entire zipping in Drosophila embryo
超分子
名称: Tomographic reconstruction of an entire zipping in Drosophila embryo タイプ: sample / ID: 1000 詳細: Joined 34 serial tomograms were reconstructed from 2x2 montaged tilt-series. Tilt-series were recorded at x12000 corresponding to 0.9nm/pixel at the specimen level. The map was binned for ...詳細: Joined 34 serial tomograms were reconstructed from 2x2 montaged tilt-series. Tilt-series were recorded at x12000 corresponding to 0.9nm/pixel at the specimen level. The map was binned for deposition to reduce the file size. The z-axis of the map corresponds to the anterior/posterior embryonic axis while the y-axis to the ventral/dorsal axis. The dorsal epidermal cells are located on the top. Number unique components: 1
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超分子 #1: zipping in Drosphila embryogenesis
超分子
名称: zipping in Drosphila embryogenesis / タイプ: organelle_or_cellular_component / ID: 1 / 組換発現: No / データベース: NCBI
由来(天然)
生物種: Drosophila melanogaster (キイロショウジョウバエ) 別称: fruit fly
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実験情報
-
構造解析
手法
ネガティブ染色法, クライオ電子顕微鏡法
解析
電子線トモグラフィー法
試料の集合状態
tissue
-
試料調製
染色
タイプ: NEGATIVE 詳細: Sections were stained with 2% UA in 70% methanol and Reynolds lead citrate.
グリッド
詳細: slot grids with Formvar support
凍結
凍結剤: NITROGEN / 装置: OTHER
-
電子顕微鏡法
顕微鏡
FEI TECNAI F30
日付
2010年5月10日
撮影
カテゴリ: FILM / フィルム・検出器のモデル: FEI EAGLE (4k x 4k) / デジタル化 - スキャナー: OTHER
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
照射モード: OTHER / 撮影モード: BRIGHT FIELD / 倍率(公称値): 12000
試料ステージ
試料ホルダーモデル: SIDE ENTRY, EUCENTRIC / Tilt series - Axis1 - Min angle: -60 ° / Tilt series - Axis1 - Max angle: 60 ° / Tilt series - Axis1 - Angle increment: 1.5 °