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Role of Mitochondrial Activation in PACAP Dependent Neurite Outgrowth

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Abstract

Pituitary adenylate cyclase-activating polypeptide (PACAP) increases neurite outgrowth, although signaling via its receptor PACAP-specific receptor (PAC1R) has not been fully characterized. Because mitochondria also play an important role in neurite outgrowth, we examined whether mitochondria contribute to PACAP-mediated neurite outgrowth. When mouse primary hippocampal neurons and Neuro2a cells were exposed to PACAP, neurite outgrowth and the mitochondrial membrane potential increased in both cell types. These results were reproduced using the PAC1R-specific agonist maxadilan and the adenylate cyclase activator forskolin, whereas the protein kinase A inhibitor H89 and mitochondrial uncoupling agent carbonyl cyanide m-chlorophenyl hydrazone (CCCP) inhibited these effects. Expression levels of peroxisome proliferator-activated receptor γ coactivator 1α (Pgc1α), a master regulator of mitochondrial activation, and its downstream effectors, such as cytochrome C and cytochrome C oxidase subunit 4, increased in response to PACAP. Knocking down Pgc1α expression using small interfering RNA or treatment with CCCP significantly attenuated neurite outgrowth and reduced the mitochondrial membrane potential in PACAP-treated cells. These data suggest that mitochondrial activation plays a key role in PACAP-induced neurite outgrowth via a signaling pathway that includes PAC1R, PKA, and Pgc1α.

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Abbreviations

ANOVA:

Analysis of variance

ATP:

Adenosine triphosphate

CCCP:

Carbonyl cyanide m-chlorophenyl hydrazone

COX4:

Cytochrome C oxidase subunit 4

CREB:

cAMP response element binding protein

CytC:

Cytochrome C

DIV:

Days in vitro

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

JC-1:

5,5′,6,6′-tetra-chloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide

MAP2:

Microtubule-associated protein 2

Δψ m :

Mitochondrial membrane potential

PBS:

Phosphate-buffered saline

PACAP:

Pituitary adenylate cyclase-activating polypeptide

PAC1R:

PACAP-specific receptor

Pgc1α:

Peroxisome proliferator-activated receptor γ coactivator 1α

PKA:

Protein kinase A

siRNA:

Small interfering RNA

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Acknowledgments

This work was supported by a Grant-in-Aid for Young Scientists (B) (22790256) from The Ministry of Education, Culture, Sports, Science and Technology (MEXT, Japan). We wish to thank Drs. Takashi Kurihara and Kazuhiko Inoue for the helpful discussions and the Joint Research Laboratory (Kagoshima University Graduate School of Medical and Dental Sciences) for the use of facilities and equipment.

Conflicts of Interest

The authors have no conflicts of interest.

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  1. Department of Pharmacology, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagosima-shi, Kagoshima, 890-8544, Japan

    Yuki Kambe & Atsuro Miyata

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  1. Yuki Kambe
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  2. Atsuro Miyata
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Correspondence to Atsuro Miyata.

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Kambe, Y., Miyata, A. Role of Mitochondrial Activation in PACAP Dependent Neurite Outgrowth. J Mol Neurosci 48, 550–557 (2012). https://doi.org/10.1007/s12031-012-9754-0

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