Journal of Molecular Neuroscience

, Volume 42, Issue 3, pp 349–358 | Cite as

Activation of STAT3 by Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) During PACAP-Promoted Neurite Outgrowth of PC12 Cells

  • Masami IshidoEmail author


Addition of pituitary adenylate cyclase-activating polypeptide (PACAP) into the cultured PC12 cells promoted neurite outgrowth of the cells, indicating cell differentiation. Using DNA macroarray techniques, we have characterized the PC12 cell transcriptome, revealing that several genes were regulated by PACAP. Among many, we focused to investigate whether STAT3 molecule, whose message was up-regulated, might be involved in PACAP signaling. Reverse transcription polymerase chain reaction supported the results obtained by DNA macroarray; PACAP increased gene expression of STAT3, at least up to 24 h, being a maximum 9.5-fold with 3 h-treatment of 1 nM PACAP. Reporter assay analyses demonstrated that PACAP activated STAT3-response promoter, but neither GAS- nor ISRE-response promoters. Enzyme-linked immunosorbent assay revealed that PACAP increased the amount of STAT3 proteins about 30%. Concurrently, phospho-STAT3 Tyr705 was increased in the nuclei by the neuropeptide. Ectopic expression of dominant negative forms of STAT3, which had Tyr705 to phenylalanine substitution, effectively decreased PACAP-promoted neurite extension (55%). During the activation of STAT3, the secretion of interleukin-6 (IL-6) was stimulated by PACAP in a dose-dependent manner. Treatment with IL-6-neutralizing antibody significantly diminished PACAP-activated STAT3 promoter activity and PACAP-induced neurite outgrowth. Thus, our findings have demonstrated that STAT3 is involved in PACAP signaling and function during PC12 cell differentiation by the neuropeptide.


PACAP STAT3 Neurite outgrowth PC12 cells 



Brain-derived neurotrophic factor


Cyclic AMP


Enzyme-linked immunosorbent assay


Extracellular regulated kinase


Interferon-gamma activation sequence


Green fluorescent protein


Glyceraldehyde-3-phosphate dehydrogenase




Interferon-stimulated response element


Mitogen-activated protein kinase


Mitogen-activated protein kinase kinase


Nerve growth factor


Phosphate-buffered saline


Pheochromocytoma cells 12


Protein kinase A


Reverse transcription polymerase chain reaction


Signal transducer and activator of transcription


TATA box of the herpes simplex virus thymus kinase promoter




Vasoactive intestinal polypeptide.



We thank Drs. K. Nakashima (Nara Institute of Science and Technology, Nara, Japan) and S. Akira (Osaka University, Osaka, Japan) for providing the expression vectors.


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Environmental Risk Research ProgrammeNational Institute for Environmental StudiesTsukubaJapan

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