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Differential distribution of α‐tubulin isotypes in Euplotes eurystomus determined by confocal immunofluorescence microscopy **
Author(s) -
Olins Donald E.,
Olins Ada L.,
RobertNicoud Michel,
Jovin Thomas M.,
Wehland Juergen,
Weber Klaus
Publication year - 1989
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1111/j.1768-322x.1989.tb00840.x
Subject(s) - biology , confocal microscopy , immunofluorescence , tubulin , confocal , differential (mechanical device) , distribution (mathematics) , microbiology and biotechnology , microtubule , antibody , immunology , optics , physics , mathematical analysis , mathematics , thermodynamics
Detergent permeabilized Euplotes eurystomus (a fresh water hypotrichous ciliate) was reacted with monoclonal and polyclonal antibodies specific for either detyrosinated or tyrosinated α‐tubulin (Glu‐ or Tyr‐tubulin). The isolated cytoskeleton‐nuclear complex was examined by Western immunoblotting and by immunofluorescent and electron microscopic methods. Both Glu‐ and Tyr‐tubulins were detected by immunoblot analysis. Immunofluorescent microscopy indicated that the α‐tubulin isotypes are concentrated in different regions of permeabilized cells: Glu‐tubulin is located primarily in cirri, membranelles, and surrounding the macro‐ and micronuclei. Tyr‐tubulin is principally at the bases of cirri and membranelles. This differential distribution of α‐tubulin isotypes is discussed in terms of current concepts concerning the correlation of tubulin post‐translational modifications to microtubule stability. Confocal immunofluorescent imaging was of critical importance in clearly differentiating the Glu‐tubulin isotype surrounding the macro‐ and micronuclei from a brilliantly fluorescent environment originating from cytoskeletal structures. In conjunction with conventional and stereo‐electron microscopy, confocal optical microscopy provided convincing evidence for a “basket” of microtubules surrounding both nuclei.