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Open Access Highly Accessed Research article

proNGF/NGF mixtures induce gene expression changes in PC12 cells that neither singly produces

Ivan Arisi1, Mara D’Onofrio12, Rossella Brandi1, Francesca Malerba35, Francesca Paoletti3, Andrea Ennio Storti1, Fulvio Florenzano4, Luisa Fasulo3 and Antonino Cattaneo35*

Author Affiliations

1 Genomics Facility, European Brain Research Institute (EBRI) “Rita Levi-Montalcini”, Via del Fosso di Fiorano, 64, 00143 Roma, Italy

2 Istituto di Farmacologia Traslazionale, CNR, Via del Fosso del Cavaliere 100, 00133 Roma, Italy

3 Neurotrophic Factors and Neurodegenerative Diseases Laboratory, European Brain Research Institute (EBRI) “Rita Levi-Montalcini”, Via del Fosso di Fiorano, 64, 00143 Roma, Italy

4 European Brain Research Institute (EBRI) “Rita Levi-Montalcini”, Confocal Microscopy Unit, Via del Fosso di Fiorano, 64, 00143 Roma, Italy

5 Scuola Normale Superiore, Piazza dei Cavalieri 7, 56126 Pisa, Italy

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BMC Neuroscience 2014, 15:48  doi:10.1186/1471-2202-15-48

Published: 8 April 2014

Abstract

Background

Growing evidence shows that, in vivo, the precursor of Nerve Growth Factor (NGF), proNGF, displays biological activities different from those of its mature NGF counterpart, mediated by distinct, and somewhat complementary, receptor binding properties. NGF and proNGF induce distinct transcriptional signatures in target cells, highlighting their different bioactivities. In vivo, proNGF and mature NGF coexist. It was proposed that the relative proNGF/NGF ratio is important for their biological outcomes, especially in pathological conditions, since proNGF, the principal form of NGF in Central Nervous System (CNS), is increased in Alzheimer’s disease brains. These observations raise a relevant question: does proNGF, in the presence of NGF, influence the NGF transcriptional response and viceversa? In order to understand the specific proNGF effect on NGF activity, depending on the relative proNGF/NGF concentration, we investigated whether proNGF affects the pattern of well-known NGF-regulated mRNAs.

Results

To test any influence of proNGF on pure NGF expression fingerprinting, the expression level of a set of candidate genes was analysed by qReal-Time PCR in rat adrenal pheochromocytoma cell line PC12, treated with a mixture of NGF and proNGF recombinant proteins, in different stoichiometric ratios. These candidates were selected amongst a set of genes well-known as being rapidly induced by NGF treatment. We found that, when PC12 cells are treated with proNGF/NGF mixtures, a unique pattern of gene expression, which does not overlap with that deriving from treatment with either proNGF or NGF alone, is induced. The specific effect is also dependent on the stoichiometric composition of the mixture. The proNGF/NGF equimolar mixture seems to partially neutralize the specific effects of the proNGF or NGF individual treatments, showing a weaker overall response, compared to the individual contributions of NGF and proNGF alone.

Conclusions

Using gene expression as a functional read-out, our data demonstrate that the relative availability of NGF and proNGF in vivo might modulate the biological outcome of these ligands.

Keywords:
PC12; NGF; proNGF; Neurotrophin; Early genes; Gene expression; Transcription; NGF/proNGF ratio