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The requirement of extracellular signal-related protein kinase pathway in the activation of hypoxia inducible factor 1α in the developing rat brain after hypoxia–ischemia

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Abstract

Hypoxia inducible factor 1α (HIF-1α) is a key regulator of cellular oxygen homeostasis. However, the regulation of HIF-1α in neonates with hypoxia–ischemia (HI) is not clear. Under normoxic conditions, the extracellular signal-related protein kinase (ERK) pathway has been shown to be involved in the activation of HIF-1α in cell lines. Therefore, we hypothesized that the ERK pathway is involved in the activation of HIF-1α and its target genes in the developing rat brain following HI. To test this hypothesis, we set up an HI model by ligating the right common carotid artery followed by hypoxia using postnatal day 10 rats. Rat brains from HI and sham controls were collected to detect the expression of HIF-1α, its target gene, vascular endothelial growth factor (VEGF), and ERK using immunohistochemistry, Western blot analysis, and RT-PCR. We found that the expression of HIF-1α protein was significantly upregulated at 4 h and peaked at 8 h after HI compared with sham controls. Accordingly, VEGF was similarly upregulated. However, the expression of total ERK (Erk1/2) had no obvious changes. Even though the phosphorylated form of ERK, p-Erk1/2, was upregulated and peaked at 4 h after HI, it is earlier than that seen in HIF-1α expression. Furthermore, the induction of HIF-1α protein, but not its mRNA, could be significantly inhibited by Erk1/2 pathway specific inhibitor, U0126. Our findings suggest that Erk1/2 pathway is involved in the regulation of HIF-1α and VEGF in the developing rat brain after HI. The Erk1/2 pathway may work as a potential target for therapeutic intervention in neonates with HI.

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Abbreviations

HIF-1α:

Hypoxia-inducible factor-1α

HI:

Hypoxia–ischemia

HIBD:

Hypoxia–ischemia brain damage

P10:

Postnatal day 10

CCA:

Common carotid artery

PBS:

Phosphate-buffered saline

PCR:

Polymerase chain reaction

Erk:

Extracellular signal-related protein kinase

MAPK:

Mitogen-active protein kinase

RT-PCR:

Reverse transcriptase-polymerase chain reaction

TBS:

Tris-buffered saline

VEGF:

Vascular endothelial growth factor

CBP:

CREB binding protein

CREB:

CAMP responsive element binding protein

References

  1. Bardos JI, Ashcroft M (2005) Negative and positive regulation of HIF-1: a complex network. Biochim Biophys Acta 1755:107–120

    PubMed  CAS  Google Scholar 

  2. Bardos JI, Ashcroft M (2004) Hypoxia-inducible factor-1 and oncogenic signaling. Bioessays 26:262–269

    Article  PubMed  CAS  Google Scholar 

  3. Conrad PW, Freeman TL, Beitner-Johnson D, Millhorn DE (1999) EPAS1 trans-activation during hypoxia requires p42/p44 MAPK. J Biol Chem 274:33709–33713

    Article  PubMed  CAS  Google Scholar 

  4. Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O’Rourke J, Mole DR, Mukherji M, Metzen E, Wilson MI, Dhanda A, Tian YM, Masson N, Hamilton DL, Jaakkola P, Barstead R, Hodgkin J, Maxwell PH, Pugh CW, Schofield CJ, Ratcliffe PJ (2001) C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation. Cell 107:43–54

    Article  PubMed  CAS  Google Scholar 

  5. Ferriero DM, Arcavi LJ, Sagar SM, McIntosh TK, Simon RP (1998) Selective sparing of NADPH-diaphorase neurons in neonatal hypoxia–ischemia. Ann Neurol 24:670–676

    Article  Google Scholar 

  6. Hagberg H, Bona E, Gilland E, Puka-Sundvall M (1997) Hypoxia–ischaemia model in the 7-day-old rat: possibilities and shortcomings. Acta Paediatr Suppl 422:85–88

    PubMed  CAS  Google Scholar 

  7. Hur E, Chang KY, Lee E, Lee SK, Park H (2001) Mitogen-activated protein kinase kinase inhibitor PD98059 blocks the trans-activation but not the stabilization or DNA binding ability of hypoxia-inducible factor-1α. Mol Pharmacol 59:1216–1224

    PubMed  CAS  Google Scholar 

  8. Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J, Gaskell SJ, Kriegsheim Av, Hebestreit HF, Mukherji M, Schofield CJ, Maxwell PH, Pugh CW, Ratcliffe PJ (2001) Targeting of HIF-1alpha to the von Hippel–Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. Science 292:468–472

    Article  PubMed  CAS  Google Scholar 

  9. Jiang BH, Jiang GQ, Zheng JZ, Lu ZM, Hunter T, Vogt PK (2001) Phosphatidylinositol 3-kinase signaling controls levels of hypoxia-inducible factor. Cell Growth Differ 12:363–369

    PubMed  CAS  Google Scholar 

  10. Jiang BH, Zheng JZ, Aoki M, Vogt PK (2000) Phosphatidylinositol 3-kinase signaling mediates angiogenesis and expression of vascular endothelial growth factor in endothelial cells. Proc Natl Acad Sci USA 97:1749–1753

    Article  PubMed  CAS  Google Scholar 

  11. Lee E, Yim S, Lee SK, Park H (2002) Two transactivation domains of hypoxia-inducible factor-1alpha regulated by the MEK-1/p42/p44 MAPK pathway. Mol Cell 14:9–15

    CAS  Google Scholar 

  12. Li L, Qu Y, Li J, Xiong Y, Mao M, Mu D (2007) Relationship between HIF-1α expression and neuronal apoptosis in neonatal rats with hypoxia–ischemia brain injury. Brain Res 1180:133–139

    Article  PubMed  CAS  Google Scholar 

  13. Mao M, Hua Y, Jiang X, Li L, Zhang L, Mu D (2006) Expression of tumor necrosis factor α and neuronal apoptosis in the developing rat brain after neonatal stroke. Neurosci Lett 403:227–232

    Article  PubMed  CAS  Google Scholar 

  14. Minet E, Michel G, Mottet D, Raes M, Michiels C (2001) Transactivation pathways involved in hypoxia-inducible factor-1 phosphorylation and activation. Free Radic Biol Med 31:847–855

    Article  PubMed  CAS  Google Scholar 

  15. Minet E, Arnould G, Michel I, Roland D, Mottet M, Raes J, Remacle R Michiels C (2000) ERK activation upon hypoxia: involvement in HIF-1 activation. FEBS Lett 468:53–58

    Article  PubMed  CAS  Google Scholar 

  16. Mottet D, Michel G, Renard P, Ninane N, Rase M, Michiels C (2002) ERK and calcium in activation of HIF-1. Ann N Y Acad Sci 973:448–453

    PubMed  CAS  Google Scholar 

  17. Mu D, Jiang X, Sheldon RA, Fox CK, Hamrick SE, Vexler ZS, Ferriero DM (2003) Regulation of hypoxia-inducible factor 1alpha and induction of vascular endothelial growth factor in a rat neonatal stroke model. Neurobiol Dis 14:524–534

    Article  PubMed  CAS  Google Scholar 

  18. Mu D, Chang YS, Vexler ZS, Ferriero DM (2005) Hypoxia-inducible factor 1alpha and erythropoietin upregulation with deferoxamine salvage after neonatal stroke. Exp Neurol 195:407–415

    Article  PubMed  CAS  Google Scholar 

  19. Richard DE, Berra E, Gothie E, Roux D, Pouyssegur J (1999) p42/p44 mitogen-activated protein kinases phosphorylate hypoxia-inducible factor 1alpha (HIF-1α) and enhance the transcriptional activity of HIF-1. J Biol Chem 274:32631–32637

    Article  PubMed  CAS  Google Scholar 

  20. Sang N, Stiehl DP, Bohensky J, Leshchinsky I, Srinivas V, Caro J (2003) MAPK signaling up-regulates the activity of hypoxia-inducible factors by its effects on p300. J Biol Chem 278:14013–14019

    Article  PubMed  CAS  Google Scholar 

  21. Semenza GL (2000) Expression of hypoxia-inducible factor 1: mechanisms and consequences. Biochem Pharmacol 59:47–53

    Article  PubMed  CAS  Google Scholar 

  22. Semenza GL (2000) Surviving ischemia: adaptive responses mediated by hypoxia-inducible factor 1. J Clin Invest 106:809–812

    Article  PubMed  CAS  Google Scholar 

  23. Senmenza GL (2000) HIF-1: using two hands to flip the angiogenic switch. Cancer Metastasis Rev 19:59–65

    Article  Google Scholar 

  24. Semenza GL (1999) Regulation of mammalian O2 homeostasis by hypoxia-inducible factor 1. Annu Rev Cell Dev Biol 15:551–578

    Article  PubMed  CAS  Google Scholar 

  25. van den Tweel ER, Kavelaars A, Lonbardi MS, Nijboer CH, Groememdaal F, Van Bel F, Heijnen CJ 2006 Bilateral molecular changes in a neonatal rat model of unilateral hypoxia–ischemia brain damage. Pediatr Res 59:434–439

    Article  PubMed  CAS  Google Scholar 

  26. Vo N, Goodman RH (2001) CREB-binding protein and p300 in transcriptional regulation. J Biol Chem 276:13505–13508

    PubMed  CAS  Google Scholar 

  27. Zheng XR, Yang YJ, Jia YJ (2003) The study of relationship between HIF-1α and neuronal apoptosis in neonatal rat brains with hypoxia–ischemia. Zhonghua Er Ke Za Zhi 41:366–367

    PubMed  Google Scholar 

  28. Zhong H, Chiles K, Feldser D, Laughner E, Hanrahan C, Georgescu MM, Simons JW, Semenza GL (2000) Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells: implications for tumor angiogenesis and therapeutics. Cancer Res 60:1541–1545

    PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by grants from National Natural Science Foundation of China (No. 30570623 and No. 30770748 to Dezhi Mu), China Medical Board of New York (00-722 to Dezhi Mu), Ministry of Education of China (2006331-11-7 and 20070610092 to Dezhi Mu), and Science and Technology Bureau of Sichuan province (JY029-067). We sincerely appreciate Stephanie Cambier from the University of California, San Francisco, for proofreading the manuscript.

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Correspondence to Dezhi Mu.

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L. Li and Y. Xiong contributed equally to this report.

This work was supported by grants from the National Natural Science Foundation of China (No. 30570623 and No. 30770748 to Dezhi Mu), China Medical Board of New York (00-722 to Dezhi Mu), Ministry of Education of China (2006331-11-7, 20070610092), and Bureau of Scientific Technology of Sichuan Province (07JY029-067).

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Li, L., Xiong, Y., Qu, Y. et al. The requirement of extracellular signal-related protein kinase pathway in the activation of hypoxia inducible factor 1α in the developing rat brain after hypoxia–ischemia. Acta Neuropathol 115, 297–303 (2008). https://doi.org/10.1007/s00401-008-0339-5

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