Uniform Polarity Microtubule Assemblies Imaged in Native Brain Tissue by Second-Harmonic Generation Microscopy

Microtubule (MT) ensemble polarity is a diagnostic determinant of the structure and function of neuronal processes. Here, polarized MT structures are selectively imaged with second-harmonic generation (SHG) microscopy in native brain tissue. This SHG is found to colocalize with axons in both brain s...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 100; no. 12; pp. 7081 - 7086
Main Authors Dombeck, Daniel A., Kasischke, Karl A., Vishwasrao, Harshad D., Ingelsson, Martin, Hyman, Bradley T., Webb, Watt W.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 10.06.2003
National Acad Sciences
Subjects
Animals
Axons
Biological Sciences
Biophysical Phenomena
Biophysics
Brain
Brain - metabolism
Brain - ultrastructure
Charge separation
Dendrites
Fluorescence
Hippocampus - metabolism
Hippocampus - ultrastructure
Imaging
Immunohistochemistry
Kinetics
Lasers
Microscopy
Microscopy, Fluorescence - methods
Microtubule-Associated Proteins - metabolism
Microtubules - metabolism
Microtubules - ultrastructure
Molecular Motor Proteins - metabolism
Nerve Tissue Proteins - metabolism
Neurons
Photons
Rats
Rats, Sprague-Dawley
tau Proteins - metabolism
Tissues
Tubulin - metabolism
Wave excitation
Wavelengths
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Summary:Microtubule (MT) ensemble polarity is a diagnostic determinant of the structure and function of neuronal processes. Here, polarized MT structures are selectively imaged with second-harmonic generation (SHG) microscopy in native brain tissue. This SHG is found to colocalize with axons in both brain slices and cultured neurons. Because SHG arises only from noninversion symmetric structures, the uniform polarity of axonal MTs leads to the observed signal, whereas the mixed polarity in dendrites leads to destructive interference. SHG imaging provides a tool to investigate the kinetics and function of MT ensemble polarity in dynamic native brain tissue structures and other subcellular motility structures based on polarized MTs.
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To whom correspondence should be addressed. E-mail: www2@cornell.edu.
Abbreviations: MT, microtubule; SH, second harmonic; SHG, SH generation; TPF, two-photon fluorescence; NA, numerical aperture; PMT, photo-multiplier tube; DG, dentate gyrus; , average excitation intensity; w, two-photon radial beam waist; MAP, MT-associated protein.
Contributed by Watt W. Webb, April 4, 2003
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0731953100