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Translational Stroke Research

, Volume 7, Issue 2, pp 132–140 | Cite as

Acute Blockage of Notch Signaling by DAPT Induces Neuroprotection and Neurogenesis in the Neonatal Rat Brain After Stroke

  • Zhongxia Li
  • Jiangping Wang
  • Congying Zhao
  • Keming Ren
  • Zhezhi Xia
  • Huimin Yu
  • Kewen JiangEmail author
Original Article

Abstract

Notch signaling is critically involved in various biological events. Notch undergoes cleavage by the γ-secretase enzyme to release Notch intracellular domain that will translocate into nucleus to result in expression of target gene. γ-Secretase inhibitors have been developed as potential treatments for neurological degenerative diseases, but its effects against ischemic injury remain relatively uncertain. In the present study, we demonstrated that N-[N-(3, 5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT), a γ-secretase inhibitor not only rescued the cerebral hypoperfusion or ischemia neonatal rats from death, reduced apoptosis in penumbra, but also reduced brain infarct size. Furthermore, DAPT elicited some morphologic hallmarks such as neurogenesis and angiogenesis that related to the brain repair and functional recovery after stroke: increased accumulations of newborn cells in the peri-infarct region with a higher fraction of them adopting immature neuronal and glial markers instead of microglial markers on 5 days, enhanced vascular densities in penumbra at 14 days, and evident regulations of the gene profiles associated with neurogenesis in penumbral tissues. The current results suggest that DAPT is a potential neuroprotectants against ischemic injury in immature brain, and future treatment strategies such as clinical trials using γ-secretase inhibitors would be an attractive therapy for perinatal ischemia.

Keywords

N-[N-(3, 5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester Notch signaling Ischemia Neuroprotection Neurogenesis Angiogenesis 

Notes

Compliance with Ethical Standards

Funding

This study was funded by the National Natural Science Foundation of China (30471840 and 30570644) and by Zhejiang Provincial Natural Science Foundation of China (Y205127 and R2090266).

Conflict of Interest

The authors declare that they have no competing interest.

Ethical Approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Supplementary material

12975_2015_441_MOESM1_ESM.doc (34 kb)
ESM 1 (DOC 34 kb)
12975_2015_441_Fig7_ESM.jpg (767 kb)
Figure S1

N-[N-(3, 5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT) regulated the gene profiles associated with neurogenesis in penumbra at 5 days after focal cerebral ischemia. The Oligo GEArray DNA Microarray Rat Neurogenesis and Neural Stem Cell Gene Array containing 263 known genes were used in the experiments. Microarray protocols were as shown in Fig. 1b. a Representative gene arrays show the profiles of genes in non-stroke, stroke, or DAPT-treated stroke penumbral tissues. All clones with more than 1.5-fold estimated differences were considered for further evaluation. Red and blue circles indicate upregulated and downregulated genes, respectively, detected in microarrays under DAPT-treated conditions compared with genes under vehicle-treated ischemia condition in the penumbra. Real-time RT-PCR analysis of mRNA levels in the penumbra. Some upregulated and downregulated genes (b) detected in microarrays under DAPT-treated condition after stroke compared with levels in vehicle-treated stroke penumbra. Quantification of expression levels of upregulated and downregulated genes (c). Values are mean ± SEM; *P < 0.05, ANOVA, n = 4. (JPG 767 kb)

12975_2015_441_Fig8_ESM.jpg (256 kb)
Figure S1

N-[N-(3, 5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT) regulated the gene profiles associated with neurogenesis in penumbra at 5 days after focal cerebral ischemia. The Oligo GEArray DNA Microarray Rat Neurogenesis and Neural Stem Cell Gene Array containing 263 known genes were used in the experiments. Microarray protocols were as shown in Fig. 1b. a Representative gene arrays show the profiles of genes in non-stroke, stroke, or DAPT-treated stroke penumbral tissues. All clones with more than 1.5-fold estimated differences were considered for further evaluation. Red and blue circles indicate upregulated and downregulated genes, respectively, detected in microarrays under DAPT-treated conditions compared with genes under vehicle-treated ischemia condition in the penumbra. Real-time RT-PCR analysis of mRNA levels in the penumbra. Some upregulated and downregulated genes (b) detected in microarrays under DAPT-treated condition after stroke compared with levels in vehicle-treated stroke penumbra. Quantification of expression levels of upregulated and downregulated genes (c). Values are mean ± SEM; *P < 0.05, ANOVA, n = 4. (JPG 767 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Zhongxia Li
    • 1
  • Jiangping Wang
    • 2
  • Congying Zhao
    • 1
  • Keming Ren
    • 1
  • Zhezhi Xia
    • 4
  • Huimin Yu
    • 3
  • Kewen Jiang
    • 1
    • 4
    Email author
  1. 1.Department of LaboratoryThe Children’s Hospital Zhejiang University School of MedicineHangzhouChina
  2. 2.Department of RehabilitationThe Children’s Hospital Zhejiang University School of MedicineHangzhouChina
  3. 3.Department of NeonatologyThe Children’s Hospital Zhejiang University School of MedicineHangzhouChina
  4. 4.Department of NeurologyThe Children’s Hospital Zhejiang University School of MedicineHangzhouChina

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