Shotgun proteomics implicates extracellular matrix proteins and protease systems in neuronal development induced by astrocyte cholinergic stimulation

Nadia H. Moore, Lucio G. Costa, Scott A. Shaffer, David R. Goodlett, Marina Guizzetti

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Astrocytes play an important role in neuronal development through the release of soluble factors that affect neuronal maturation. Shotgun proteomics followed by gene ontology analysis was used in this study to identify proteins present in the conditioned medium of primary rat astrocytes. One hundred and thirty three secreted proteins were identified, the majority of which were never before reported to be produced by astrocytes. Extracellular proteins were classified based on their biological and molecular functions; most of the identified proteins were involved in neuronal development. Semi-quantitative proteomic analysis was carried out to identify changes in the levels of proteins released by astrocytes after stimulation with the cholinergic agonist carbachol, as we have previously reported that carbachol-treated astrocytes elicit neuritogenesis in hippocampal neurons through the release of soluble factors. Carbachol up-regulated secretion of 15 proteins and down-regulated the release of 17 proteins. Changes in the levels of four proteins involved in neuronal differentiation (thrombospondin-1, fibronectin, plasminogen activator inhibitor-1, and plasminogen activator urokinase) were verified by western blot or ELISA. In conclusion, this study identified a large number of proteins involved in neuronal development in the astrocyte secretome and implicated extracellular matrix proteins and protease systems in neuronal development induced by astrocyte cholinergic stimulation.

Original languageEnglish (US)
Pages (from-to)891-908
Number of pages18
JournalJournal of Neurochemistry
Volume108
Issue number4
DOIs
StatePublished - Feb 2009
Externally publishedYes

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Extracellular Matrix Proteins
Firearms
Astrocytes
Proteomics
Cholinergic Agents
Peptide Hydrolases
Proteins
Carbachol
Thrombospondin 1
Cholinergic Agonists
Gene Ontology
Plasminogen Activators
Plasminogen Activator Inhibitor 1
Urokinase-Type Plasminogen Activator
Conditioned Culture Medium
Fibronectins
Neurons
Ontology
Rats
Genes

Keywords

  • Astrocyte-released proteins
  • Astrocytes
  • Muscarinic receptors
  • Shotgun proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Shotgun proteomics implicates extracellular matrix proteins and protease systems in neuronal development induced by astrocyte cholinergic stimulation. / Moore, Nadia H.; Costa, Lucio G.; Shaffer, Scott A.; Goodlett, David R.; Guizzetti, Marina.

In: Journal of Neurochemistry, Vol. 108, No. 4, 02.2009, p. 891-908.

Research output: Contribution to journalArticle

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