Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures

Adrienne M Benediktsson; Scott J Schachtele; Steven H Green; Michael E Dailey

doi:10.1016/j.jneumeth.2004.05.013

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Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures

Journal article

Peer reviewed

Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures

Adrienne M Benediktsson, Scott J Schachtele, Steven H Green and Michael E Dailey

Journal of neuroscience methods, Vol.141(1), pp.41-53

2005

DOI: 10.1016/j.jneumeth.2004.05.013

PMID: 15585287

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Abstract

Protoplasmic astrocytes in mammalian CNS tissues in vivo have a highly complex 3D morphology, but in dissociated cell cultures they often assume a flattened, fibroblast-like morphology bearing only a few, simple processes. By fluorescent labeling and confocal reconstruction we show that many astrocytes in organotypic hippocampal slice cultures exhibit a more native complex cytoarchitecture. Although astrocytes at the surface of slice cultures show a reactive form with several thick glial fibrillary acidic protein (GFAP)-positive processes, astrocytes situated in deeper portions of tissue slices retain a highly complex 3D morphology with many fine spine- or veil-like protrusions. Dozens of astrocytes can be labeled in single slice cultures by gene gun-mediated ballistic delivery of gold or tungsten particles carrying cDNAs (Biolistics), lipophilic dyes (DiOlistics), or fluorescent intracellular calcium indicators (Calistics). Expression of a membrane-targeted form of eGFP (Lck-GFP) is superior to soluble eGFP for resolving fine astrocytic processes. Time-lapse confocal imaging of Lck-GFP transfected astrocytes or “calistically” labeled astrocytes show structural remodeling and calcium transients, respectively. This approach provides an in vitro system for investigating the functional architecture, development and dynamic remodeling of astrocytes and their relationships to neurons and glia in live mammalian brain tissues.

GFP

Calcium

Time-lapse

Morphology

Biolistics

Confocal microscopy

Astrocyte

Slice culture

Details

Title: Subtitle: Ballistic labeling and dynamic imaging of astrocytes in organotypic hippocampal slice cultures
Creators: Adrienne M Benediktsson
Scott J Schachtele
Steven H Green
Michael E Dailey
Resource Type: Journal article
Publication Details: Journal of neuroscience methods, Vol.141(1), pp.41-53
Publisher: Elsevier B.V
DOI: 10.1016/j.jneumeth.2004.05.013
PMID: 15585287
ISSN: 0165-0270
eISSN: 1872-678X
Language: English
Date published: 2005
Academic Unit: Iowa Neuroscience Institute; Biology; Otolaryngology
Record Identifier: 9984070676802771

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