Analysis of Dendritic Spine Morphology in Cultured CNS Neurons

Deepak P. Srivastava; Kevin M. Woolfrey; Peter Penzes

doi:10.3791/2794

Analysis of Dendritic Spine Morphology in Cultured CNS Neurons

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Analysis of Dendritic Spine Morphology in Cultured CNS Neurons

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11:48 min

July 13, 2011

DOI: 10.3791/2794-v

Deepak P. Srivastava¹, Kevin M. Woolfrey¹, Peter Penzes^1,2

¹Department of Physiology,Northwestern University Feinberg School of Medicine, ²Department of Psychiatry and Behavioral Sciences,Northwestern University Feinberg School of Medicine

Overview

This study investigates the effects of molecular and pharmacological manipulations on dendritic spine morphology and motility in primary cultured cortical neurons. By utilizing GFP and DNA plasmids, researchers can visualize and manipulate neuronal structures and functions.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Pharmacology

Background

Mutations in synaptic proteins are linked to brain pathologies.
Primary cultured cortical neurons are valuable for studying neuronal morphology.
Dendritic spines are critical for synaptic function and plasticity.
Pharmacological treatments can induce changes in spine morphology.

Purpose of Study

To assess the impact of various manipulations on dendritic spine characteristics.
To visualize changes in spine morphology and motility.
To understand the role of specific proteins in dendritic spine dynamics.

Methods Used

Transection of primary cultured cortical neurons with GFP.
Exogenous expression of DNA plasmids to manipulate protein expression.
Pharmacological treatment of cells for visualization.
Immunostaining and time-lapse imaging using confocal microscopy.

Main Results

Detailed measurements of dendritic spine morphology were obtained.
Changes in spine motility were observed following pharmacological treatments.
Visualization techniques provided insights into neuronal structure dynamics.
Results contribute to understanding synaptic alterations in brain pathologies.

Conclusions

The study highlights the importance of dendritic spine morphology in neuronal function.
Pharmacological manipulations can effectively alter spine characteristics.
Findings may inform future research on synaptic dysfunction in diseases.

Frequently Asked Questions

What are dendritic spines?

Dendritic spines are small protrusions on neurons that are sites of synaptic connections and play a key role in synaptic plasticity.

How does pharmacological treatment affect dendritic spines?

Pharmacological treatment can induce changes in the morphology and motility of dendritic spines, impacting synaptic function.

What is the significance of using primary cultured cortical neurons?

Primary cultured cortical neurons provide a controlled environment to study neuronal behavior and the effects of specific manipulations.

What imaging techniques are used in this study?

Confocal microscopy is used for both fixed and live cell imaging to visualize dendritic spines.

What role do synaptic proteins play in brain pathologies?

Mutations in synaptic proteins can disrupt normal spine function and contribute to various neurological disorders.

Numerous recent studies have identified mutations in synaptic proteins associated with brain pathologies. Primary cultured cortical neurons offer great flexibility in examining the effects of these disease-associated proteins on dendritic spine morphology and motility.

The overall goal of the following experiment is to assess the effects of various molecular and pharmacological manipulations on dendritic spine morphology and motility. This is achieved by transecting primary cultured cortical neurons with GFP, with or without exogenous expression of DNA plasmids to outline neuronal morphology and to manipulate endogenous protein expression and function. In the second step, cells can be treated pharmacologically and subsequently fixed into immuno stain or prepared for time-lapse imaging.

And then pharmacological treatment, which will allow for the visualization of dendritic spines and the induced changes in dendritic spine morphology and motility. Next Z Series images of fixed cells or images of live cells taken at specific time points are acquired using a confocal microscope. Detailed measurements of dendritic spine morphology can be made from 2D projections of Z series images.

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