Automated 3D quantitative analysis of dendritic spines imaged with light microscopy

Abstract This project aims to develop a novel systemSpines InvestigatorTMfor performing automated four dimensionalDquantitative analysis of changes in dendritic spine morphology on three dimensionalDmicroscopic images acquired with in vivo multiphoton fluorescence microscopy at different time pointsThe role of dendritic spines is one of the most active and important areas of neuroscience researchPlasticity of dendritic spine morphology plays a crucial role throughout lifein developmentagingas well as in learning and memoryAlsomany complex brain diseasesincluding autism spectrum disordersschizophreniaDown syndromeAlzheimerandapos s diseaseParkinsonandapos s diseaseHuntingtonandapos s diseaseamyotrophic lateral sclerosis and strokeare characterized by dendritic spine pathology including abnormal dendritic spine density and morphologydendritic spine lossand aberrant dendritic spine plasticityWhile it is not possible to study the plasticity of dendritic spine morphology in the human brain in vivoit is possible in mouse models of complex human brain diseasesHoweverthe study of these mouse models remains a tedious and cumbersome endeavor because tools for automatedD dendritic spine quantitative analysis are not availableCritical steps that are currently performed manually in such investigations may lead to faulty and irreproducible resultswhich does not conform with NIHandapos s rigor and transparency policySpines Investigator will help solve this untenable situation with a number of distinct innovationsSpecificallySpines Investigator will comprise novel technology that enables the automated comparison of dendritic spine morphology onD images acquired with in vivo multiphoton fluorescence microscopy in the brain of a mouse at precisely the same site at different time pointsIt will also enable new research that combinesD in vivo quantitative analysis of changes in dendritic spine morphology with the analysis of amyloid plaquesin Alzheimerandapos s diseaseas well as analysis of microglia and astrocytesTo create this new solution for automatedD in vivo quantitative analysis of dendritic spine morphologySpines Investigator will build upon our Neurolucidatechnology developed during Phase IISBIR Fast track Grant MHWe will develop Spines Investigator as a testedvalidatedsupported and fully documented systemThe benefit for the neuroscience research communitypharmacological and biotechnological research and developmentand society in general will be to better understand the critical role of the plasticity of dendritic spine morphology in the brain under various physiological and pathological conditionsIn particularthis will result in an improved basis for developing novel treatment strategies for complex brain diseases Narrative Analysis of the plasticity and dynamics of dendritic spine morphology in living animals with in vivo multiphoton fluorescence microscopy yields important information about nervous system function in brain development and aginglearning and memoryand the dysfunction of the brain in many psychiatricneurological and childhood developmental disordersThis project creates a much needed innovative system for automated quantitative analysis of the changes in dendritic spine morphology on three dimensional microscopic images acquired in the brain of a mouse at the same site at different time pointsmaking such analyses for the first time fully comply with NIHandapos s rigor and transparency policyThis will result in an improved basis for developing novel treatment strategies for complex CNS diseases such as autism spectrum disordersRett syndromeschizophreniaFragile X syndromeDown syndromeAlzheimerandapos s diseaseParkinsonandapos s diseaseHuntingtonandapos s diseaseamyotrophic lateral sclerosisstroke and many other brain diseases and conditions