4.3 Article

Automated Sholl Analysis of Digitized Neuronal Morphology at Multiple Scales: Whole Cell Sholl Analysis Versus Sholl Analysis of Arbor Subregions

Journal

CYTOMETRY PART A
Volume 77A, Issue 12, Pages 1160-1168

Publisher

WILEY
DOI: 10.1002/cyto.a.20954

Keywords

Sholl analysis; neurite; BDNF; morphology; computer-assisted; digitization; tracing

Funding

  1. Busch Biomedical Grant
  2. NSF [IBN-0548543, IBN-0919747, DGE-0333196]
  3. March of Dimes Foundation [1-FY04-107, 1-FY08-464]
  4. NIH [T32 GM008339-20, 5 T32 MH019957-10, T32 GM00839]
  5. NJ Commission [08-2941-SCR-E-0]
  6. University and Louis Bevier Graduate and Dissertation Fellowship
  7. IGERT at Rutgers
  8. Division Of Integrative Organismal Systems
  9. Direct For Biological Sciences [0919747] Funding Source: National Science Foundation

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The morphology of dendrites and the axon determines how a neuron processes and transmits information. Neurite morphology is frequently analyzed by Sholl analysis or by counting the total number of neurites and branch tips. However, the time and resources required to perform such analysis by hand is prohibitive for the processing of large data sets and introduces problems with data auditing and reproducibility. Furthermore, analyses performed by hand or using course-grained morphometric data extraction tools can obscure subtle differences in data sets because they do not store the data in a form that facilitates the application of multiple analytical tools. To address these shortcomings, we have developed a program (titled Bonfire) to facilitate digitization of neurite morphology and subsequent Sholl analysis. Our program builds upon other available open-source morphological analysis tools by performing Sholl analysis on subregions of the neuritic arbor, enabling the detection of local level changes in dendrite and axon branching behavior. To validate this new tool, we applied Bonfire analysis to images of hippocampal neurons treated with 25 ng/ml brain-derived neurotrophic factor (BDNF) and untreated control neurons. Consistent with prior findings, conventional Sholl analysis revealed that global exposure to BDNF increases the number of neuritic intersections proximal to the soma. Bonfire analysis additionally uncovers that BDNF treatment affects both root processes and terminal processes with no effect on intermediate neurites. Taken together, our data suggest that global exposure of hippocampal neurons to BDNF results in a reorganization of neuritic segments within their arbors, but not necessarily a change in their number or length. These findings were only made possible by the neurite-specific Sholl data returned by Bonfire analysis. (C) 2010 International Society for Advancement of Cytometry

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