Determination of microtubule polarity in vitro by the use of video-enhanced differential-interference contrast light microscopy and Chlamydomonas flagellar axonemal pieces. 1995

T C Gamblin, and R C Williams
Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235, USA.

Microtubules nucleated by sea urchin sperm-tail axonemes have polar ends that differ both functionally and structurally but cannot be distinguished from one another when viewed by light microscopy. Ambiguity and circularity surround any classification of microtubule polarity by conventional methods. Chlamydomonas flagellar axonemal pieces have distinct morphological differences at their plus- and minus-ends, and microtubules nucleated from these pieces can be distinguished as plus- or minus-ended based on the morphological differences present in the Chlamydomonas flagellar axonemal pieces. Plus- and minus-ended microtubules were polymerized in this fashion and analyzed for differences in growth rates, shortening rates, and frequencies of transitions. The results were in good agreement with similar data generated by the more time-consuming and difficult use of kinesin-coated beads (R. J. Kowalski, and R. C. Williams, Jr. (1993) Cell Motil. Cytoskeleton 26, 282-290) to determine microtubule polarity. This is a relatively simple and effective method for determining the polarity of microtubules in vitro by video-enhanced differential-interference contrast light microscopy.

UI MeSH Term Description Entries
D008297 Male Males
D008857 Microscopy, Interference The science and application of a double-beam transmission interference microscope in which the illuminating light beam is split into two paths. One beam passes through the specimen while the other beam reflects off a reference mirror before joining and interfering with the other. The observed optical path difference between the two beams can be measured and used to discriminate minute differences in thickness and refraction of non-stained transparent specimens, such as living cells in culture. Interferometry, Microscopic,Microinterferometry,Microscopy, Differential Interference Contrast,Microscopy, Interference Reflection,Microscopy, Nomarski Interference Contrast,Interference Microscopy,Interference Reflection Microscopy,Microscopic Interferometry,Reflection Microscopy, Interference
D008863 Microspheres Small uniformly-sized spherical particles, of micrometer dimensions, frequently labeled with radioisotopes or various reagents acting as tags or markers. Latex Beads,Latex Particles,Latex Spheres,Microbeads,Bead, Latex,Beads, Latex,Latex Bead,Latex Particle,Latex Sphere,Microbead,Microsphere,Particle, Latex,Particles, Latex,Sphere, Latex,Spheres, Latex
D008870 Microtubules Slender, cylindrical filaments found in the cytoskeleton of plant and animal cells. They are composed of the protein TUBULIN and are influenced by TUBULIN MODULATORS. Microtubule
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013082 Sperm Tail The posterior filiform portion of the spermatozoon (SPERMATOZOA) that provides sperm motility. Sperm Flagellum,Flagellum, Sperm,Flagellums, Sperm,Sperm Flagellums,Sperm Tails,Tail, Sperm,Tails, Sperm
D014404 Tubulin A microtubule subunit protein found in large quantities in mammalian brain. It has also been isolated from SPERM FLAGELLUM; CILIA; and other sources. Structurally, the protein is a dimer with a molecular weight of approximately 120,000 and a sedimentation coefficient of 5.8S. It binds to COLCHICINE; VINCRISTINE; and VINBLASTINE. alpha-Tubulin,beta-Tubulin,delta-Tubulin,epsilon-Tubulin,gamma-Tubulin,alpha Tubulin,beta Tubulin,delta Tubulin,epsilon Tubulin,gamma Tubulin
D016547 Kinesins A family of microtubule-associated mechanical adenosine triphosphatases, that uses the energy of ATP hydrolysis to move organelles along microtubules including mitosis, meiosis, and axonal transport. Kinesin,Kinesin Heavy-Chain Protein,Kinesin Light-Chain Protein,Kinesin Light-Chain Proteins,Kinesin Superfamily,Heavy-Chain Protein, Kinesin,Light-Chain Protein, Kinesin,Light-Chain Proteins, Kinesin,Protein, Kinesin Heavy-Chain,Protein, Kinesin Light-Chain,Proteins, Kinesin Light-Chain,Superfamily, Kinesin
D016825 Chlamydomonas reinhardtii A species of GREEN ALGAE. Delicate, hairlike appendages arise from the flagellar surface in these organisms. Chlamydomonas reinhardii,Chlamydomonas reinhardius,Chlamydomonas reinhardtius,reinhardius, Chlamydomonas,reinhardtii, Chlamydomonas
D018715 Microscopy, Video Microscopy in which television cameras are used to brighten magnified images that are otherwise too dark to be seen with the naked eye. It is used frequently in TELEPATHOLOGY. Video Microscopy,Videomicrography,Videomicroscopy,Microscopies, Video,Video Microscopies,Videomicrographies,Videomicroscopies

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