Abstract

Genomic and proteomic studies of brain regions of specialized function provide evidence that communication among neurons is mediated by systems of diverse chemical messengers. These analyses are largely tissueor population-based, whereas the actual communication is from cell-to-cell. To understand the complement of intercellular signals produced by individual neurons, new methods are required. We have developed a novel neuron-to-neuron, serum-free coculture approach that was used to determine the higher-level cellular peptidome of individual primary mammalian neurons. We isolated magnocellular neurons from the supraoptic nucleus of early postnatal rat and maintained them in serum-free low-density cultures without glial support layers; under these conditions, they required low-density cocultured neurons. Coculturing magnocellular neurons with hippocampal neurons permitted local access to individual neurons within the culture for mass spectrometry. Using direct sampling, we obtained peptide profiles for spatially distinct, identifiable neurons within the coculture. We repeatedly detected 10 peaks that we assign to previously characterized peptides and 17 peaks that remain unassigned. Peptides from the vasopressin prohormone and secretogranin-2 are attributed to magnocellular neurons, whereas neurokinin A, peptide J, and neurokinin B are attributed to cultured hippocampal neurons. This approach enables the elucidation of cell-specific prohormone processing and the discovery of cell-cell signaling peptides.

Original languageEnglish (US)
Pages (from-to)36-48
Number of pages13
JournalACS Chemical Neuroscience
Volume1
Issue number1
DOIs
StatePublished - Jan 20 2010

Fingerprint

Coculture Techniques
Neurons
Peptides
Neurokinin B
Chromogranins
Cell signaling
Neurokinin A
Supraoptic Nucleus
Communication
Serum
Vasopressins
Cell Communication
Neuroglia
Proteomics
Mass spectrometry
Rats
Brain
Mass Spectrometry
Sampling

Keywords

  • Cell culture
  • Hippocampus
  • Magnocellular neuron
  • Mass spectrometry
  • Neuron coculture
  • Neuropeptide
  • Supraoptic nucleus

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Physiology
  • Cognitive Neuroscience

Cite this

Direct cellular peptidomics of supraoptic magnocellular and hippocampal neurons in low-density cocultures. / Millet, Larry J.; Bora, Adriana; Sweedler, Jonathan V.; Gillette, Martha U.

In: ACS Chemical Neuroscience, Vol. 1, No. 1, 20.01.2010, p. 36-48.

Research output: Contribution to journalArticle

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