Differential labelling of trypanosome microtubules using tubulin subunit monoclonal antibodies

Cell Biology

2

International

Reports,

Vol. 8, No.

7, January

1984

DIFFERENTIAL LABELLING OF TRYPANOSOMEMICROTUBULESUSING TUBULIN SUBUNIT MONOCLONALANTIEODIES R.Cumning and J.Williamson National

Institute

for

Medical

Research,

Mill

Hill,

London NW7 1AA

Our previous studies in brain have shown that two nonoclonal antibodies YLl/Z and YOL/* recognising different regions of the a tubulin subunit, detect different popylations of axonal nicrotubules .In the present study we have used these antibodies(and one against B tubulin) for innuno fluorescent studies in Trypanosona rhodesiense.This parasitic protozoon is also of interest since a recent study using the closely re lated --T.brucei has used tubulin nonoclonal antibodies to show differential staining of the flagellar and2sub-pellicular microtubules . Blood smears from rats infected with a monomorphic strain of T. rhodesiense were fixed in cold acetone and stained by indirect innunofluoresence;the subunit/is0 type characterisation of the a tubulin antibodies(raised agai s yeast tubulin) is described in f3'. Our results show that while anti body YOL/$(a tubulin) and the @ tubulin antibody label both flagellar and cell body(sub-pellicular > nicrotubules,staining with YL1/2 (a tubulin) is restricted to the flagella and bright spots ie. the basal bodies(red blood cells are unstained). Antibody YL1/2 has recently been shown to etect only tyrosylated CI tubulin 3 .Previous work has shown that the different microtubule in a unicellular organism. systems in trypanosones nay have& Acknowledgements: Dr J.Kilmartin varying biochemical compositions and Mr.T.Scott-Finnigan. and s ow different sensitivity to l.Eur,J.Cell Bioi,3!-,2&l.-2?;r3 taxol 3 .The present results suggest 2.EMBO J. 2,479-'483 that immunocytochenistry can label 3.5 Cell Biol In press (1983) different forms of c( tubulin in iC.Biochen. 17,2883-2891 func-tionally different structures 5.Proc Nat1 Acad Sci 78,4571-4575 Received:

1st November 1383.

0309-1651/84/010002-01~$03.00/0

Accepted: @ 1984

14th November 1983.

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Inc. (London)

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