3-DIMENSIONAL STRUCTURAL CHARACTERIZATION OF CENTROSOMES FROM EARLY DROSOPHILA EMBRYOS

被引:183
作者
MORITZ, M [1 ]
BRAUNFELD, MB [1 ]
FUNG, JC [1 ]
SEDAT, JW [1 ]
ALBERTS, BM [1 ]
AGARD, DA [1 ]
机构
[1] UNIV CALIF SAN FRANCISCO, HOWARD HUGHES MED INST, SAN FRANCISCO, CA 94143 USA
关键词
D O I
10.1083/jcb.130.5.1149
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
An understanding of the mechanism and structure of microtubule (MT)-nucleating sites within the pericentriolar material (PCM) of the centrosome has been elusive. This is partly due to the difficulty in obtaining large quantities of centrosomes for analysis, as well as to the problem of attaining interpretable structural data with conventional EM techniques, We describe a protocol for isolating a large quantity of functional centrosomes from early Drosophila embryos. Using automated electron tomography, we have begun a three-dimensional structural characterization of these intact centrosomes with and without regrown MTs. Reconstructions of the centrosomes to similar to 6-8 nm resolution revealed no large structures at the minus ends of MTs, suggesting that if MT-nucleating material physically contacts the MTs, it must conform closely to the shape of the minus end. While many MTs originate near the centrioles, MT minus ends were found throughout the PCM, and even close to its outer boundary. The MTs criss-crossed the PCM, suggesting that nucleating sites are oriented in many different directions. Reconstructions of centrosomes without MTs suggest that there is a reorganization of the PCM upon MT regrowth; moreover, ring-like structures that have a similar diameter as MTs are apparent in the PCM of centrosomes without MTs, and may be MT-nucleating sites.
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页码:1149 / 1159
页数:11
相关论文
共 57 条
[1]  
Alieva I. B., 1992, P103
[2]  
ASHBURNER M, 1989, DROSOPHILA LABORATOR
[3]   TENSION, MICROTUBULE REARRANGEMENTS, AND THE PROPER DISTRIBUTION OF CHROMOSOMES IN MITOSIS [J].
AULT, JG ;
NICKLAS, RB .
CHROMOSOMA, 1989, 98 (01) :33-39
[4]   A 3-DIMENSIONAL APPROACH TO MITOTIC CHROMOSOME STRUCTURE - EVIDENCE FOR A COMPLEX HIERARCHICAL ORGANIZATION [J].
BELMONT, AS ;
SEDAT, JW ;
AGARD, DA .
JOURNAL OF CELL BIOLOGY, 1987, 105 (01) :77-92
[5]   REAL-TIME VISUALIZATION OF CELL-CYCLE DEPENDENT CHANGES IN MICROTUBULE DYNAMICS IN CYTOPLASMIC EXTRACTS [J].
BELMONT, LD ;
HYMAN, AA ;
SAWIN, KE ;
MITCHISON, TJ .
CELL, 1990, 62 (03) :579-589
[6]   POLARITY OF MICROTUBULES NUCLEATED BY CENTROSOMES AND CHROMOSOMES OF CHINESE-HAMSTER OVARY CELLS-INVITRO [J].
BERGEN, LG ;
KURIYAMA, R ;
BORISY, GG .
JOURNAL OF CELL BIOLOGY, 1980, 84 (01) :151-159
[7]  
BOZZOLA JJ, 1992, ELECTRON MICROS, P390
[8]   CRYO AUTOMATED ELECTRON TOMOGRAPHY - TOWARDS HIGH-RESOLUTION RECONSTRUCTIONS OF PLASTIC-EMBEDDED STRUCTURES [J].
BRAUNFELD, MB ;
KOSTER, AJ ;
SEDAT, JW ;
AGARD, DA .
JOURNAL OF MICROSCOPY, 1994, 174 :75-84
[9]   TUBULIN ASSEMBLY SITES AND THE ORGANIZATION OF CYTOPLASMIC MICROTUBULES IN CULTURED MAMMALIAN-CELLS [J].
BRINKLEY, BR ;
COX, SM ;
PEPPER, DA ;
WIBLE, L ;
BRENNER, SL ;
PARDUE, RL .
JOURNAL OF CELL BIOLOGY, 1981, 90 (03) :554-562
[10]  
BYERS B, 1978, J CELL SCI, V30, P331