Membrane targeted horseradish peroxidase as a marker for correlative fluorescence and electron microscopy studies

Li, J., Wang, Y., Chiu, S. L., Cline, H. T. (2010) Membrane targeted horseradish peroxidase as a marker for correlative fluorescence and electron microscopy studies. Front Neural Circuits, 4. p. 6. ISSN 1662-5110

URL: http://www.ncbi.nlm.nih.gov/pubmed/20204144
DOI: 10.3389/neuro.04.006.2010

Abstract

Synaptic dynamics and reorganization are fundamental features of synaptic plasticity both during synaptic circuit development and in the mature CNS underlying learning, memory, and experience-dependent circuit rearrangements. Combining in vivo time-lapse fluorescence imaging and retrospective electron microscopic analysis provides a powerful technique to decipher the rules governing dynamics of neuronal structure and synaptic connections. Here we have generated a membrane-targeted horseradish peroxidase (mHRP) that allows identification of transfected cells without obscuring the intracellular ultrastructure or organelles and in particular allows identification of synaptic sites using electron microscopy. The expression of mHRP does not affect dendritic arbor growth or dynamics of transfected neurons. Co-expression of EGFP and mHRP was used to study neuronal morphology at both the light and electron microscopic levels. mHRP expression greatly facilitates 3D reconstruction based on serial EM sections. We expect this reagent will be valuable for studying the mechanisms that guide construction of neuronal networks.

Item Type: Paper
Uncontrolled Keywords: 3D reconstruction Xenopus laevis horseradish peroxidase synapse time-lapse imaging ultrastructure
Subjects: Investigative techniques and equipment > microscopy > electron microscopy
Investigative techniques and equipment > microscopy > immunoflourescence microscopy
CSHL Authors:
Communities: CSHL labs > Cline lab
Watson School > Publications
Depositing User: Matt Covey
Date: 2010
Date Deposited: 22 Sep 2014 19:23
Last Modified: 22 Sep 2014 19:23
PMCID: PMC2831632
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30799

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving