Preparation of synaptoneurosomes from mouse cortex using a discontinuous percoll-sucrose density gradient

Pamela R. Westmark, Cara J. Westmark, Athavi Jeevananthan, James S. Malter

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Synaptoneurosomes (SNs) are obtained after homogenization and fractionation of mouse brain cortex. They are resealed vesicles or isolated terminals that break away from axon terminals when the cortical tissue is homogenized. The SNs retain pre- and postsynaptic characteristics, which makes them useful in the study of synaptic transmission. They retain the molecular machinery used in neuronal signaling and are capable of uptake, storage, and release of neurotransmitters.The production and isolation of active SNs can be problematic using medias like Ficoll, which can be cytotoxic and require extended centrifugation due to high density, and filtration and centrifugation methods, which can result in low activity due to mechanical damage of the SNs. However, the use of discontinuous Percoll-sucrose density gradients to isolate SNs provides a rapid method to produce good yields of translationally active SNs. The Percoll-sucrose gradient method is quick and gentle as it employs isotonic conditions, has fewer and shorter centrifugation spins and avoids centrifugation steps that pellet SNs and cause mechanical damage.

Original languageEnglish (US)
Article numbere3196
JournalJournal of Visualized Experiments
Issue number55
DOIs
StatePublished - Sep 2011

Fingerprint

Centrifugation
Sugar (sucrose)
Sucrose
Ficoll
Gradient methods
Presynaptic Terminals
Fractionation
Synaptic Transmission
Machinery
Neurotransmitter Agents
Brain
Tissue
Percoll

Keywords

  • Cortex
  • Issue 55
  • Mouse
  • Neurons
  • Neuroscience
  • Percoll-sucrose gradients
  • Synapse
  • Synaptoneurosomes
  • Synaptosomes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Preparation of synaptoneurosomes from mouse cortex using a discontinuous percoll-sucrose density gradient. / Westmark, Pamela R.; Westmark, Cara J.; Jeevananthan, Athavi; Malter, James S.

In: Journal of Visualized Experiments, No. 55, e3196, 09.2011.

Research output: Contribution to journalArticle

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