Centrin and the cytoskeleton of the protist Holomastigotoides

Wilma L. Lingle, Jeffrey L Salisbury

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The cytoskeleton of the parabasalid protozoan Holomastigotoides was investigated by epifluorescence, scanning confocal, and transmission electron microscopy using antibodies to centrin, tubulin, and MPM-2 epitopes. Previous microscopic analysis of Holomastigotoides spp. has shown that up to 10,000 flagella are arranged in 2-8 spiral bands encircling the cell. Spindle poles are associated with two flagellar bands. Sheets of cytoplasmic microtubules (MTs) called axostyles originate in the cell apex and extend to the cell base. Antibodies to centrin, a member of the EF-hand family of calcium- binding proteins, labeled a number of structures in Holomastigotoides, namely axostyles, the mitotic spindle, and portions of flagellar bands. The identity of these structures was confirmed by transmission electron microscopy and by immunofluorescence microscopy using antibodies to tubulin and MPM-2 epitopes. Antibodies to tubulin labeled MTs in basal bodies, flagella, axostyles, and the mitotic spindle. MPM-2 antibodies labeled spindle poles and short segments of flagellar bands to which the spindle poles are attached. Centrin is known to show calcium-sensitive contractile behavior. The pattern of flagellar band staining by antibodies to centrin was affected by [Ca2+]. In detergent-extracted cell fragments, the centrin staining pattern could be changed by changing [Ca2+]. This Ca2+ effect was modulated by a monoclonal antibody to centrin (20H5), indicating that centrin plays a role in altering flagellar band structure. These results show that centrin is located in key positions for maintaining cell polarity and directing cell movement through interactions with other cytoskeletal elements. Calcium may regulate the morphology of centrin-containing structures.

Original languageEnglish (US)
Pages (from-to)377-390
Number of pages14
JournalCell Motility and the Cytoskeleton
Volume36
Issue number4
DOIs
StatePublished - 1997

Fingerprint

Sulfamethoxazole Drug Combination Trimethoprim
Cytoskeleton
Spindle Poles
Antibodies
Tubulin
Spindle Apparatus
Flagella
Microtubules
Parabasalidea
Epitopes
Staining and Labeling
Basal Bodies
Calcium
EF Hand Motifs
Cell Polarity
Scanning Transmission Electron Microscopy
Calcium-Binding Proteins
Transmission Electron Microscopy
Fluorescence Microscopy
Detergents

Keywords

  • basal bodies
  • fibrillar structures
  • flagellar bands
  • immunofluorescence microscopy

ASJC Scopus subject areas

  • Cell Biology

Cite this

Centrin and the cytoskeleton of the protist Holomastigotoides. / Lingle, Wilma L.; Salisbury, Jeffrey L.

In: Cell Motility and the Cytoskeleton, Vol. 36, No. 4, 1997, p. 377-390.

Research output: Contribution to journalArticle

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