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Microglia-derived proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1beta induce Purkinje neuronal apoptosis via their receptors in hypoxic neonatal rat brain

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Abstract

The developing cerebellum is extremely vulnerable to hypoxia which can damage the Purkinje neurons. We hypothesized that this might be mediated by tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) derived from activated microglia as in other brain areas. One-day-old rats were subjected to hypoxia following, which the expression changes of various proteins in the cerebellum including hypoxia inducible factor-1α, TNF-α, IL-1β, TNF-R1 and IL-1R1 were analyzed. Following hypoxic exposure, TNF-α and IL-1β immunoexpression in microglia was enhanced coupled by that of TNF-R1 and IL-1R1 in the Purkinje neurons. Along with this, hypoxic microglia in vitro showed enhanced release of TNF-α and IL-1β whose receptor expression was concomitantly increased in the Purkinje neurons. In addition, nitric oxide (NO) level was significantly increased in the cerebellum and cultured microglia subjected to hypoxic exposure. Moreover, cultured Purkinje neurons treated with conditioned medium derived from hypoxic microglia underwent apoptosis but the incidence was significantly reduced when the cells were treated with the same medium that was neutralized with TNF-α/IL-1β antibody. We conclude that hypoxic microglia in the neonatal cerebellum produce increased amounts of NO, TNF-α and IL-1β which when acting via their respective receptors could induce Purkinje neuron death.

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Abbreviations

BSA:

Bovine serum albumin

DAPI:

4′-6-Diamidino-2-phenylindole

DMEM:

Dulbecco’s modified Eagle’s medium

ELISA:

Enzyme-linked immunosorbent assay

HIF-1α:

Hypoxia inducible factor-1α

iNOS:

Inducible nitric oxide synthase

IL-1β:

Interleukin-1β

IL-1R1 :

Interleukin-1 receptor 1

NO:

Nitric oxide

PBS:

Phosphate buffered saline

PVDF:

Polyvinylidene difluoride

TNF-α:

Tumor necrosis factor-α

TNF-R1 :

Tumor necrosis factor receptor 1

TUNEL:

Terminal deoxynucleotidyl transferase (Tdt)-mediated dUTP nick end labeling

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Acknowledgments

This study was supported by a research grant (R181-000-120-213) from National Medical Research Council of Singapore. Z. Zou was supported by a grant (No. 81260297) from the National Sciences Foundation of China.

Conflict of interest

There is no conflict of interest among the authors.

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Authors and Affiliations

  1. Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, Blk MD10, 4 Medical Drive, Singapore, 117597, Singapore

    Charanjit Kaur, Viswanathan Sivakumar & Eng-Ang Ling

  2. Department of Anatomy, Faculty of Basic Medical Sciences, Kunming Medical University, 1168 West Chunrong Road, Kunming, 650500, People’s Republic of China

    Zhirong Zou

Authors
  1. Charanjit Kaur
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  2. Viswanathan Sivakumar
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  3. Zhirong Zou
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  4. Eng-Ang Ling
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Correspondence to Charanjit Kaur.

Additional information

C. Kaur and E.-A. Ling contributed equally to the project.

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Kaur, C., Sivakumar, V., Zou, Z. et al. Microglia-derived proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1beta induce Purkinje neuronal apoptosis via their receptors in hypoxic neonatal rat brain. Brain Struct Funct 219, 151–170 (2014). https://doi.org/10.1007/s00429-012-0491-5

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