Polyclonal antibodies against human gamma-tubulin stain centrioles in mammalian cells from different tissues.
Rabbit polyclonal antibodies were raised against the C-terminal fragment (amino acid residues 318-451) of human gamma-tubulin. These antibodies were used to stain cultured cells of various tissues (epithelium, nervous tissue, fibroblasts) from different animals (human, monkey, pig, rat, kangaroo rat, mouse, hamster, chicken, triton). The antibodies specifically stained centrioles in the interphase and mitotic cells of mammals, but not birds (chicken) or amphibians (newt). In the interphase cells, centrioles were stained as a pair of dots (or as a double dot) in 96-97% of the cells. The distances between the maternal and filial centrioles varied in different cultures. Procentrioles were stained in certain cells, but with less intensity than mature centrioles. In mitotic cells, the antibodies revealed two spots corresponding to two mitotic poles. The spots in mitosis were significantly larger than the interphase dots, but the staining was more faint. In spontaneous tripolar mitoses, only two poles were stained. Thus, it was shown that, on the one hand, gamma-tubulin is associated with centrioles irrespective of whether or not they serve as the microtubule organizing centres and, on the other hand, gamma-tubulin might not be an essential component of the microtubule organizing centres.
Komarova YuA
Belozersky Institute of Physico-Chemical Biology, Moscow State University.
Ryabov
E V
EV
Alieva
I B
IB
Uzbekov
R E
RE
Uzbekova
S V
SV
Vorobjev
L A
LA
D000818:Animals; D000906:Antibodies; D002478:Cells, Cultured; D002502:Centrioles / Q000378:metabolism*; D002645:Chickens; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D007399:Interphase / Q000502:physiology; D007614:Macropodidae; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis / Q000502:physiology; D009417:Nerve Tissue / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D012455:Salamandridae; D013194:Staining and Labeling / Q000379:methods; D013552:Swine; D014404:Tubulin / Q000276:immunology* / Q000378:metabolism*; D014709:Vero Cells
D000906:Antibodies; D014404:Tubulin