Optimally both spines and neurons should be examined at high resolution, but frequently a compromise is necessary for throughput. It is also important to assess entire neurons or dendritic trees as it permits to assess tree complexity and dendritic branching, both of which may be altered in disease states. Therefore, there is value in imaging and quantifying specific spine populations, types, densities, and distribution along neuronal dendrites across brain regions or cortical layers.
Developmental, aging-, and disease-related structural changes in neurons and dendritic spines, and their functional consequences, remain poorly understood. Spine morphology determines the strength, stability and function of excitatory synaptic connections that subserve the neuronal networks underlying cognitive function.
These protocols will be a valuable reference for scientists working towards quantifying and characterizing spines.īecause of their capacity for plasticity and their role as the location of excitatory synapses, accurately characterizing the structure of dendritic spines is of profound biological significance. This unit presents methods for objective dendritic spine morphometric analysis by providing image acquisition parameters needed to ensure optimal data series for proper spine detection, characterization, and quantification with Neurolucida 360. While there have been some automated software packages that quantify spine number, they are limited in terms of their characterization of spine structure. However, this quantification has been restricted to time- and labor-intensive methods such as electron microscopy and manual counting, which have limited throughput and are impractical for studies of large samples. Precise quantification of spines in three dimensions (3D) is essential for understanding the structural determinants of normal and pathological neuronal function.
Determining the density and morphology of dendritic spines is of high biological significance given the role of spines in synaptic plasticity and in neurodegenerative and neuropsychiatric disorders.
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