Semi-automated Sholl analysis for quantifying changes in growth and differentiation of neurons and glia

John C. Gensel, David L. Schonberg, Jessica K. Alexander, Dana M. McTigue, Phillip G. Popovich

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

There is a need to develop therapies that promote growth or remyelination of mammalian CNS axons. Although the feasibility of pre-clinical treatment strategies should be tested in animal models, in vitro assays are usually faster and less expensive. As a result, in vitro models are ideal for screening large numbers of potential therapeutics prior to use in more complex in vivo systems. In 1953, Sholl introduced a technique that is a reliable and sensitive method for quantifying indices of neurite outgrowth. However, application of the technique is limited because it is labor-intensive. Several methods have been developed to reduce the analysis time for the Sholl technique; but these methods require extensive pre-processing of digital images, they introduce user bias or they have not been compared to manual analysis to ensure accuracy. Here we describe a new, semi-automated Sholl technique for quantifying neuronal and glial process morphology. Using MetaMorph®, we developed an unbiased analysis protocol that can be performed ∼3× faster than manual quantification with a comparable level of accuracy regardless of cell morphology. The laborious image processing typical of most computer-aided analysis is avoided by embedding image correction functions into the automated portion of the analysis. The sensitivity and validity of the technique was confirmed by quantifying neuron growth treated with growth factors or oligodendroglial maturation in the presence or absence of thyroid hormone. Thus, this technique provides a rapid and sensitive method for quantifying changes in cell morphology and screening for treatment effects in multiple cell types in vitro.

Original languageEnglish
Pages (from-to)71-79
Number of pages9
JournalJournal of Neuroscience Methods
Volume190
Issue number1
DOIs
StatePublished - Jun 2010

Bibliographical note

Funding Information:
This work is supported by The Craig H. Neilsen Foundation; Paralysis Project of America; NINDS NS037846 and NS059776. We are grateful to Dr. Daniel Ankeny and Dr. Satoshi Nakamura for their assistance in developing Sholl analysis in our laboratory and to Akshata Almad for help generating OPC cultures and labeling oligodendrocyte lineage cells. We also thank Dr. Kristina Kigerl and Dr. Ming Wang for technical assistance with DRG preparations and Dr. Metin Gurcan for advice about the experimental design.

Keywords

  • BDNF
  • Dorsal root ganglion
  • Morphometric
  • Morphometry
  • Myelin
  • NGF
  • Oligodendrocyte progenitor
  • Regeneration
  • Sensory
  • Technique

ASJC Scopus subject areas

  • Neuroscience (all)

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