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dc.contributor.author
Shahmoradian, Sarah H.
dc.contributor.author
Galiano, Mauricio Raul
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dc.contributor.author
Wu, Chengbiao
dc.contributor.author
Chen, Shurui
dc.contributor.author
Rasband, Matthew N.
dc.contributor.author
Mobley, William C.
dc.contributor.author
Chiu, Wah
dc.date.available
2021-04-20T18:31:23Z
dc.date.issued
2014-02
dc.identifier.citation
Shahmoradian, Sarah H.; Galiano, Mauricio Raul; Wu, Chengbiao; Chen, Shurui; Rasband, Matthew N.; et al.; Preparation of primary neurons for visualizing neurites in a frozen-hydrated state using cryo-electron tomography; MyJoVE Corp.; Journal of Visualized Experiments; 84; e50783; 2-2014; 1-12
dc.identifier.issn
1940-087X
dc.identifier.uri
http://hdl.handle.net/11336/130530
dc.description.abstract
Neurites, both dendrites and axons, are neuronal cellular processes that enable the conduction of electrical impulses between neurons. Defining the structure of neurites is critical to understanding how these processes move materials and signals that support synaptic communication. Electron microscopy (EM) has been traditionally used to assess the ultrastructural features within neurites; however, the exposure to organic solvent during dehydration and resin embedding can distort structures. An important unmet goal is the formulation of procedures that allow for structural evaluations not impacted by such artifacts. Here, we have established a detailed and reproducible protocol for growing and flash-freezing whole neurites of different primary neurons on electron microscopy grids followed by their examination with cryo-electron tomography (cryo-ET). This technique allows for 3-D visualization of frozen, hydrated neurites at nanometer resolution, facilitating assessment of their morphological differences. Our protocol yields an unprecedented view of dorsal root ganglion (DRG) neurites, and a visualization of hippocampal neurites in their near-native state. As such, these methods create a foundation for future studies on neurites of both normal neurons and those impacted by neurological disorders.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
MyJoVE Corp.
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
BRAIN
dc.subject
CRYO-ELECTRON MICROSCOPY
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ELECTRON MICROSCOPE TOMOGRAPHY
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ISSUE 84
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MORPHOLOGICAL ASSAY
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NEURONS
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NEUROSCIENCE
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PRIMARY NEURON CULTURE
dc.subject
RAT
dc.subject.classification
Bioquímica y Biología Molecular
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Preparation of primary neurons for visualizing neurites in a frozen-hydrated state using cryo-electron tomography
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2021-03-26T19:37:51Z
dc.journal.volume
84
dc.journal.number
e50783
dc.journal.pagination
1-12
dc.journal.pais
Estados Unidos
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dc.journal.ciudad
Cambridge
dc.description.fil
Fil: Shahmoradian, Sarah H.. Baylor University; Estados Unidos
dc.description.fil
Fil: Galiano, Mauricio Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina. Baylor University; Estados Unidos
dc.description.fil
Fil: Wu, Chengbiao. Baylor University; Estados Unidos
dc.description.fil
Fil: Chen, Shurui. Baylor University; Estados Unidos
dc.description.fil
Fil: Rasband, Matthew N.. Baylor University; Estados Unidos
dc.description.fil
Fil: Mobley, William C.. Baylor University; Estados Unidos
dc.description.fil
Fil: Chiu, Wah. Baylor University; Estados Unidos
dc.journal.title
Journal of Visualized Experiments
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.jove.com/10.3791/50783
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.3791/50783
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