Vibrodissociation of neurons from rodent brain slices to study synaptic transmission and image presynaptic terminals.

Sang Beom Jun, Verginia Cuzon Carlson, Stephen Ikeda, David Lovinger

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Mechanical dissociation of neurons from the central nervous system has the advantage that presynaptic boutons remain attached to the isolated neuron of interest. This allows for examination of synaptic transmission under conditions where the extracellular and postsynaptic intracellular environments can be well controlled. A vibration-based technique without the use of proteases, known as vibrodissociation, is the most popular technique for mechanical isolation. A micropipette, with the tip fire-polished to the shape of a small ball, is placed into a brain slice made from a P1-P21 rodent. The micropipette is vibrated parallel to the slice surface and lowered through the slice thickness resulting in the liberation of isolated neurons. The isolated neurons are ready for study within a few minutes of vibrodissociation. This technique has advantages over the use of primary neuronal cultures, brain slices and enzymatically isolated neurons including: rapid production of viable, relatively mature neurons suitable for electrophysiological and imaging studies; superior control of the extracellular environment free from the influence of neighboring cells; suitability for well-controlled pharmacological experiments using rapid drug application and total cell superfusion; and improved space-clamp in whole-cell recordings relative to neurons in slice or cell culture preparations. This preparation can be used to examine synaptic physiology, pharmacology, modulation and plasticity. Real-time imaging of both pre- and postsynaptic elements in the living cells and boutons is also possible using vibrodissociated neurons. Characterization of the molecular constituents of pre- and postsynaptic elements can also be achieved with immunological and imaging-based approaches.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number51
StatePublished - 2011
Externally publishedYes

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Presynaptic Terminals
Synaptic Transmission
Neurons
Rodentia
Brain
Imaging techniques
Pharmacology
Physiology
Clamping devices
Neurology
Patch-Clamp Techniques
Vibration
Cell culture
Plasticity
Fires
Peptide Hydrolases
Central Nervous System
Cell Culture Techniques
Cells
Modulation

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Vibrodissociation of neurons from rodent brain slices to study synaptic transmission and image presynaptic terminals. / Jun, Sang Beom; Cuzon Carlson, Verginia; Ikeda, Stephen; Lovinger, David.

In: Journal of visualized experiments : JoVE, No. 51, 2011.

Research output: Contribution to journalArticle

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