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Mitogen-Activated Protein Kinase Pathways are Involved in the Upregulation of Calcitonin Gene-Related Peptide of Rat Trigeminal Ganglion After Organ Culture

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Abstract

The trigeminal ganglion (TG) can express and release calcitonin gene-related peptide (CGRP), an important neuropeptide that plays a crucial role in migraine attack and cluster headache. Activation of rat TG increases CGRP expression. However, the regulatory mechanism of CGRP expression in TG neurons remains to be explored. This study aims to evaluate the involvement of mitogen-activated protein kinase (MAPK) pathways in CGRP upregulation after rat TG organ culture. Rat TG was cultured alone for 24 h or cultured in combination with MAPK inhibitors, tumor necrosis factor α (TNF-α), or interleukin 1β (IL-1β) for 24 h. CGRP protein was determined using immunohistochemistry. The mRNA levels of CGRP, TNF-α, and IL-1β were analyzed through real-time quantitative polymerase chain reaction. MAPK phosphorylation was detected via western blot. After rat TG organ culture, the expressions of CGRP, TNF-α, and IL-1β were upregulated at 24 h. The phosphorylation of extracellular signal-regulated kinases (ERK1/2), P38, and c-jun N-terminal kinases (JNK) significantly increased at 30 min compared with fresh rat TG. In addition, both CGRP expression and phosphorylation of ERK1/2, P38, and JNK were enhanced obviously after rat TG treatment with TNF-α or IL-1β compared with fresh rat TG. However, they decreased markedly after rat TG pretreatment with PD98059 (ERK1/2 inhibitor), SB203580 (P38 inhibitor), or SP600125 (JNK inhibitor) compared with rat TG co-culture with TNF-α or IL-1β. In conclusion, the elevated CGRP expression after rat TG organ culture can be regulated via MAPK pathways. The findings provide insight into the molecular mechanisms and experimental evidence for therapeutic targets of migraine.

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Abbreviations

CGRP:

Calcitonin gene-related peptide

ERK1/2:

Extracellular signal-regulated kinases 1/2

IL-1β:

Interleukin 1β

JNK:

c-Jun N-terminal kinases

MAPK:

Mitogen-activated protein kinases

P38:

P38 protein

TG:

Trigeminal ganglion

TNF-α:

Tumor necrosis factor α

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Acknowledgments

We should express much gratitude to Mr. Fujun Zhang, Ms. Yan Han, and Qilan Ning for their expertise and assistance in the experiments. This study is supported by the National Natural Science Foundation of China (grant no. 30570631, 30630058).

Conflict of interest

There were no conflicts of interest in relation to this article.

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Authors and Affiliations

  1. Department of Genetics and Molecular Biology, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi’an Jiaotong University College of Medicine, Xi’an, Shaanxi, 710061, People’s Republic of China

    Li Lei, Shaolan Wang, Fujun Zhang, Yan Han, Qilan Ning & Shemin Lu

  2. Department of Neurology, The First Affiliated Hospital of Xi’an Jiaotong University College of Medicine, Xi’an, Shaanxi, 710061, People’s Republic of China

    Xingyun Yuan & Guogang Luo

  3. Department of Epidemiology and Medical Statistics, School of Public Health, Xi’an Jiaotong University College of Medicine, Xi’an, Shaanxi, 710061, People’s Republic of China

    Shemin Lu

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  1. Li Lei
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  2. Xingyun Yuan
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  3. Shaolan Wang
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  4. Fujun Zhang
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  5. Yan Han
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  6. Qilan Ning
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  7. Guogang Luo
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  8. Shemin Lu
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Correspondence to Guogang Luo or Shemin Lu.

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Lei, L., Yuan, X., Wang, S. et al. Mitogen-Activated Protein Kinase Pathways are Involved in the Upregulation of Calcitonin Gene-Related Peptide of Rat Trigeminal Ganglion After Organ Culture. J Mol Neurosci 48, 53–65 (2012). https://doi.org/10.1007/s12031-012-9772-y

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