Classification of axonal subtypes based on cytoskeletal components

Ye Z. Spector, Qi Zhao, Xiaopeng Zhao, William J Feuer, Portia Lynn Maravich, XiangRun Huang

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background: Retinal ganglion cells are often classified into different subtypes according to their morphology or physiological functions. The axons of RGCs contain three major cytoskeletal components: actin filaments (F-actin); microtubules; and neurofilaments (NFs). The contents of these components vary among axons. Our objective was to classify axons into subtypes based on the contents of cytoskeletal components and study their distributions across the retina in normal rodent retinas. Methods: Whole-mounted retinas of female Wistar rats were stained with phalloidin to label F-actin, anti-β-tubulin monoclonal antibody to mark microtubules, and antineurofilament antibody to label NFs. A confocal laser scanning microscope was used to provide en face images of retinal nerve fiber bundles with a resolution of 0.24 μm/pixel. Staining intensity profiles across axons were obtained for each cytoskeletal component. Axonal subtypes were then determined from the relative contents, indicated by the staining intensity, of these components. Linear density was used to investigate topographical distribution of each subtype across the retina. Results: Normal axons could be classified into seven subtypes - FMN, FM, FN, and MN subtypes, (in which at least two or three cytoskeletal components were intensely stained), and F, M, and N subtypes, (in which only one cytoskeletal component was intensely stained within an axon). The FMN subtype was the most abundant subtype. There was no preferential distribution of subtypes around the optic nerve head. However, the densities of the axonal subtypes that contained NFs were found significantly different in the central and peripheral retinal regions. Axonal sizes were subtype-dependent. Conclusion: Axons of retinal ganglion cells can be classified into different subtypes, based on the contents of axonal cytoskeletal components. The classified subtypes will provide a new means to study selective damage of axonal ultrastructures in ocular neuropathic diseases.

Original languageEnglish
Pages (from-to)1-10
Number of pages10
JournalCell Health and Cytoskeleton
Volume6
DOIs
StatePublished - Apr 11 2014

Fingerprint

Axons
Intermediate Filaments
Retina
Flavin Mononucleotide
Actins
Retinal Ganglion Cells
Microtubules
Labels
Staining and Labeling
Phalloidine
Eye Diseases
Optic Disk
Tubulin
Actin Cytoskeleton
Nerve Fibers
Wistar Rats
Rats
Rodentia
Optics
Lasers

Keywords

  • Cytoskeletal components
  • F-actin
  • Microtubules
  • Neurofilaments
  • Retina

ASJC Scopus subject areas

  • Cell Biology
  • Structural Biology
  • Histology
  • Biochemistry

Cite this

Classification of axonal subtypes based on cytoskeletal components. / Spector, Ye Z.; Zhao, Qi; Zhao, Xiaopeng; Feuer, William J; Maravich, Portia Lynn; Huang, XiangRun.

In: Cell Health and Cytoskeleton, Vol. 6, 11.04.2014, p. 1-10.

Research output: Contribution to journalArticle

Spector, Ye Z. ; Zhao, Qi ; Zhao, Xiaopeng ; Feuer, William J ; Maravich, Portia Lynn ; Huang, XiangRun. / Classification of axonal subtypes based on cytoskeletal components. In: Cell Health and Cytoskeleton. 2014 ; Vol. 6. pp. 1-10.
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abstract = "Background: Retinal ganglion cells are often classified into different subtypes according to their morphology or physiological functions. The axons of RGCs contain three major cytoskeletal components: actin filaments (F-actin); microtubules; and neurofilaments (NFs). The contents of these components vary among axons. Our objective was to classify axons into subtypes based on the contents of cytoskeletal components and study their distributions across the retina in normal rodent retinas. Methods: Whole-mounted retinas of female Wistar rats were stained with phalloidin to label F-actin, anti-β-tubulin monoclonal antibody to mark microtubules, and antineurofilament antibody to label NFs. A confocal laser scanning microscope was used to provide en face images of retinal nerve fiber bundles with a resolution of 0.24 μm/pixel. Staining intensity profiles across axons were obtained for each cytoskeletal component. Axonal subtypes were then determined from the relative contents, indicated by the staining intensity, of these components. Linear density was used to investigate topographical distribution of each subtype across the retina. Results: Normal axons could be classified into seven subtypes - FMN, FM, FN, and MN subtypes, (in which at least two or three cytoskeletal components were intensely stained), and F, M, and N subtypes, (in which only one cytoskeletal component was intensely stained within an axon). The FMN subtype was the most abundant subtype. There was no preferential distribution of subtypes around the optic nerve head. However, the densities of the axonal subtypes that contained NFs were found significantly different in the central and peripheral retinal regions. Axonal sizes were subtype-dependent. Conclusion: Axons of retinal ganglion cells can be classified into different subtypes, based on the contents of axonal cytoskeletal components. The classified subtypes will provide a new means to study selective damage of axonal ultrastructures in ocular neuropathic diseases.",
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AB - Background: Retinal ganglion cells are often classified into different subtypes according to their morphology or physiological functions. The axons of RGCs contain three major cytoskeletal components: actin filaments (F-actin); microtubules; and neurofilaments (NFs). The contents of these components vary among axons. Our objective was to classify axons into subtypes based on the contents of cytoskeletal components and study their distributions across the retina in normal rodent retinas. Methods: Whole-mounted retinas of female Wistar rats were stained with phalloidin to label F-actin, anti-β-tubulin monoclonal antibody to mark microtubules, and antineurofilament antibody to label NFs. A confocal laser scanning microscope was used to provide en face images of retinal nerve fiber bundles with a resolution of 0.24 μm/pixel. Staining intensity profiles across axons were obtained for each cytoskeletal component. Axonal subtypes were then determined from the relative contents, indicated by the staining intensity, of these components. Linear density was used to investigate topographical distribution of each subtype across the retina. Results: Normal axons could be classified into seven subtypes - FMN, FM, FN, and MN subtypes, (in which at least two or three cytoskeletal components were intensely stained), and F, M, and N subtypes, (in which only one cytoskeletal component was intensely stained within an axon). The FMN subtype was the most abundant subtype. There was no preferential distribution of subtypes around the optic nerve head. However, the densities of the axonal subtypes that contained NFs were found significantly different in the central and peripheral retinal regions. Axonal sizes were subtype-dependent. Conclusion: Axons of retinal ganglion cells can be classified into different subtypes, based on the contents of axonal cytoskeletal components. The classified subtypes will provide a new means to study selective damage of axonal ultrastructures in ocular neuropathic diseases.

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