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

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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.

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Abbreviations

nAChR:

nicotinic acetylcholine receptor

MLA:

Methyllycaconitine citrate

CCh:

Carbamoylcholine

b-FGF:

basic fibroblast growth factor

dbcAMP:

dibutyril cAMP

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Acknowledgments

The work was supported by research grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), project no.: 2006/61285-9, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Brazil, awarded to H.U.; A.A. N.'s and C.A.T.'s Ph.D. theses are supported by fellowships from FAPESP, Brazil. A.H.M and V.A.E. acknowledge the NIH grant support (UPR-PRAABREP20RR016470 and G12RR03035-24), R.R.R is grateful for grants from CNPq and FAPEMIG.

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Authors and Affiliations

  1. Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil

    Arthur A. Nery, Cleber A. Trujillo & Henning Ulrich

  2. Department of Physics, Institute of Exact Sciences, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil

    Rodrigo R. Resende

  3. Federal University of São João Del-Rei-Campus Centro-Oeste, Divinópolis-MG, Brazil

    Rodrigo R. Resende

  4. Department of Biochemistry, University Central del Caribe, Bayamon, Puerto Rico

    Antonio H. Martins & Vesna A. Eterovic

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  1. Arthur A. Nery
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  2. Rodrigo R. Resende
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  6. Henning Ulrich
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Correspondence to Henning Ulrich.

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Nery, A.A., Resende, R.R., Martins, A.H. et al. Alpha7 Nicotinic Acetylcholine Receptor Expression and Activity During Neuronal Differentiation of PC12 Pheochromocytoma Cells. J Mol Neurosci 41, 329–339 (2010). https://doi.org/10.1007/s12031-010-9369-2

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  • DOI: https://doi.org/10.1007/s12031-010-9369-2

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