Abstract
The shape, structure and connectivity of nerve cells are important aspects of neuronal function. Genetic and epigenetic factors that alter neuronal morphology or synaptic localization of pre- and post-synaptic proteins contribute significantly to neuronal output and may underlie clinical states. To assess the impact of individual genes and disease-causing mutations on neuronal morphology, reliable methods are needed. Unfortunately, manual analysis of immuno-fluorescence images of neurons to quantify neuronal shape and synapse number, size and distribution is labor-intensive, time-consuming and subject to human bias and error. We have developed an automated image analysis routine using steerable filters and deconvolutions to automatically analyze dendrite and synapse characteristics in immuno-fluorescence images. Our approach reports dendrite morphology, synapse size and number but also synaptic vesicle density and synaptic accumulation of proteins as a function of distance from the soma as consistent as expert observers while reducing analysis time considerably. In addition, the routine can be used to detect and quantify a wide range of neuronal organelles and is capable of batch analysis of a large number of images enabling high-throughput analysis.
| Original language | English |
|---|---|
| Pages (from-to) | 185-93 |
| Number of pages | 9 |
| Journal | Journal of Neuroscience Methods |
| Volume | 195 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 15 Feb 2011 |
| Externally published | Yes |
Keywords
- Animals
- Automatic Data Processing/methods
- Cells, Cultured
- Dendrites/metabolism
- Diagnostic Imaging
- Disks Large Homolog 4 Protein
- Guanylate Kinases
- Hippocampus/cytology
- Intracellular Signaling Peptides and Proteins/metabolism
- Lysine/analogs & derivatives
- Lysosome-Associated Membrane Glycoproteins/metabolism
- Membrane Proteins/metabolism
- Mice
- Mice, Mutant Strains
- Microtubule-Associated Proteins/metabolism
- Munc18 Proteins/genetics
- Neurites/metabolism
- Neurons/cytology
- Neuropeptide Y/metabolism
- Receptors, Transferrin/metabolism
- Software
- Synapses/physiology
- Synaptic Vesicles/metabolism
- Time Factors
- Vesicle-Associated Membrane Protein 2/metabolism