Journal of Molecular Neuroscience

, Volume 41, Issue 3, pp 329–339

Alpha7 Nicotinic Acetylcholine Receptor Expression and Activity During Neuronal Differentiation of PC12 Pheochromocytoma Cells

  • Arthur A. Nery
  • Rodrigo R. Resende
  • Antonio H. Martins
  • Cleber A. Trujillo
  • Vesna A. Eterovic
  • Henning Ulrich
Article

DOI: 10.1007/s12031-010-9369-2

Cite this article as:
Nery, A.A., Resende, R.R., Martins, A.H. et al. J Mol Neurosci (2010) 41: 329. doi:10.1007/s12031-010-9369-2

Abstract

Nicotinic acetylcholine receptors (nAChR) exert pivotal roles in synaptic transmission, neuroprotection and differentiation. Particularly, homomeric α7 receptors participate in neurite outgrowth, presynaptic control of neurotransmitter release and Ca2+ influx. However, the study of recombinant α7 nAChRs in transfected cell lines is difficult due to low expression of functional receptor channels. We show that PC12 pheochromocytoma cells induced to differentiation into neurons are an adequate model for studying differential nAChR gene expression and receptor activity. Whole-cell current recording indicated that receptor responses increased during the course of differentiation. Transcription of mRNAs coding for α3, α5, α7, β2 and β4 subunits was present during the course of differentiation, while mRNAs coding for α2, α4 and β3 subunits were not expressed in PC12 cells. α7 subunit expression was highest following 1 day of induction to differentiation. Activity of α7 nAChRs, however, was most elevated on day 2 as revealed by inhibition experiments in the presence of 10 nM methyllycaconitine, rapid current decay and receptor responsiveness to the α7 agonist choline. Increased α7 receptor activity was noted when PC12 were induced to differentiation in the presence of choline, confirming that chronic agonist treatment augments nAChR activity. In summary, PC12 cells are an adequate model to study the role and pharmacological properties of this receptor during neuronal differentiation.

Keywords

Nicotinic acetylcholine receptorsPC12 pheochromocytoma cellsAlpha7 subtypesWhole-cell recordingNicotinic receptor expression during differentiation

Abbreviations

nAChR

nicotinic acetylcholine receptor

MLA

Methyllycaconitine citrate

CCh

Carbamoylcholine

b-FGF

basic fibroblast growth factor

dbcAMP

dibutyril cAMP

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Arthur A. Nery
    • 1
  • Rodrigo R. Resende
    • 2
    • 3
  • Antonio H. Martins
    • 4
  • Cleber A. Trujillo
    • 1
  • Vesna A. Eterovic
    • 4
  • Henning Ulrich
    • 1
  1. 1.Departamento de Bioquímica, Instituto de QuímicaUniversidade de São PauloSão PauloBrazil
  2. 2.Department of Physics, Institute of Exact SciencesFederal University of Minas GeraisBelo HorizonteBrazil
  3. 3.Federal University of São João Del-Rei-Campus Centro-OesteDivinópolis-MGBrazil
  4. 4.Department of BiochemistryUniversity Central del CaribeBayamonPuerto Rico