Improved visualization of F-actin in the green alga Acetabularia by microwave-accelerated fixation and simultaneous FITC-Phalloidin staining

被引:9
作者
Sawitzky, H [1 ]
WillingaleTheune, J [1 ]
Menzel, D [1 ]
机构
[1] MAX PLANCK INST CELL BIOL,D-68522 LADENBURG,GERMANY
来源
HISTOCHEMICAL JOURNAL | 1996年 / 28卷 / 05期
关键词
D O I
10.1007/BF02331398
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
By employing a new procedure we have been able to visualize a highly intense actin cytoskeleton in the unicellular green alga Acetabularia acetabulum Silva. The protocol described in this study involves microwave-accelerated simultaneous permeabilization with 10% dimethyl sulphoxide, fixation with 1% glutaraldehyde and incubation with 0.5 mu M fluorescein-isothiocyanate-conjugated Phalloidin. Comparison of the images of the actin cytoskeleton of the stalk, as visualized by methods used previously, with those obtained in our own experiments shows that the actin filaments were preserved completely in an excellent condition. The required time for each procedure could be reduced from 12 h for the most commonly used immunofluorescence technique to 35 min. Moreover, it has been possible to observe the actin filament system of hair whorls, rhizoid and tip. Previously, the actin cytoskeleton of these parts of the cell could not be visualized by conventional techniques. It is shown that each region of the cell - stalk, tip, rhizoid and side branches - displays characteristic degrees of actin bundling and regularity of actin alignment.
引用
收藏
页码:353 / 360
页数:8
相关论文
共 30 条
[1]  
BERGER S, 1992, DASYCLADALES ILILLUS
[2]   MICROWAVES FOR IMMUNOHISTOCHEMISTRY [J].
BOON, ME ;
KOK, LP .
MICRON, 1994, 25 (02) :151-170
[3]   EFFECTS OF CYTOCHALASIN AND PHALLOIDIN ON ACTIN [J].
COOPER, JA .
JOURNAL OF CELL BIOLOGY, 1987, 105 (04) :1473-1478
[4]   REACTIVATION OF CYTOPLASMIC STREAMING IN A TONOPLAST-PERMEABILIZED CELL MODEL OF ACETABULARIA [J].
FUKUI, S ;
NAGAI, R .
PLANT AND CELL PHYSIOLOGY, 1985, 26 (04) :737-744
[5]  
GIBOR A, 1978, PROGR ACETABULARIA R, P231
[6]   CONTROL OF REGENERATION AND MORPHOGENESIS BY DIVALENT-CATIONS IN ACETABULARIA-MEDITERRANEA [J].
GOODWIN, BC ;
SKELTON, JL ;
KIRKBELL, SM .
PLANTA, 1983, 157 (01) :1-7
[7]   FIXATION INDUCES DIFFERENTIAL TIP MORPHOLOGY AND IMMUNOLOCALIZATION OF THE CYTOSKELETON IN POLLEN TUBES [J].
HE, Y ;
WETZSTEIN, HY .
PHYSIOLOGIA PLANTARUM, 1995, 93 (04) :757-763
[8]   CYTOCHALASIN STOPS TIP GROWTH IN PLANTS [J].
HERTH, W ;
FRANKE, WW ;
VANDERWOUDE, WJ .
NATURWISSENSCHAFTEN, 1972, 59 (01) :38-+
[9]   CYCLICAL TRANSFORMATIONS OF THE ACTIN CYTOSKELETON OF HYACINTH POLLEN SUBJECTED TO RECURRENT VAPOR-PHASE HYDRATION AND DEHYDRATION [J].
HESLOPHARRISON, J ;
HESLOPHARRISON, Y .
BIOLOGY OF THE CELL, 1992, 75 (03) :245-252
[10]  
HESLOPHARRISON J, 1991, SEX PLANT REPROD, V4, P6