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Angiogenic Gene Profiles in Laser-Microdissected Microvessels and Neurons from Ischemic Penumbra of Rat Brain

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Abstract

Angiogenesis is induced immediately after cerebral ischemia and plays a pivotal role in the strategy against ischemic injury. We hypothesized that the coordinated interaction between microvessels and neurons was altered immediately after stroke, and microvessels and neurons would show the temporal specificity of angiogenic gene profiles after cerebral ischemia. Microvessels and neurons were harvested in the ischemic penumbra of rat brain using the PixCell II laser capture microdissection (LCM) instrument. After RNA isolation, T7 and gene-specific primer RNA linear amplification were performed, and angiogenic functional grouping cDNA profiling was analyzed in LCM samples. cDNA microarray results showed there were 35 (36.46%) and 27 (28.13%) genes expression changes in the microvessels, while 25 (26.04%) and 31 (32.29%) genes were changed in the neurons at 2 h and 24 h after cerebral ischemia. Members of growth factors and receptors, cytokines and chemokines, adhesion molecules, matrix proteins, proteases, and inhibitors showed temporal and spatial differentiation in the microvessels and neurons after cerebral ischemia. This finding will help to understand the coordination and interaction between microvessels and neurons, and to elucidate the molecular mechanisms of angiogenesis after brain ischemic injury.

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Acknowledgements

We thank the Laboratory Animal Research Center at Xi’an Jiaotong College University of Medicine for their animal care and support, and Xiaoge Zhao for slice cutting.

Funding

This research was supported by the National Natural Science Foundation of China (31371501), Natural Science Foundation of Shaanxi Province (2017JM8047, 2018JM7066), and the Fundamental Research Funds for the Central Universities (GK201702006), Shaanxi Normal University.

Author information

Authors and Affiliations

  1. Key Laboratory of Modern Teaching Technology, Ministry of Education, Xi’an, 710062, Shaanxi, China

    Yingfang Tian & Meimei Shi

  2. College of Life Sciences, Shaanxi Normal University, Xi’an, 710119, Shaanxi, China

    Yingfang Tian, Yuanyuan Di, Ting Feng, Frank Adu-Nti & Juan Fan

  3. Institute of Neurobiology, School of Basic Medical Sciences, Xi’an Jiaotong University Health Science Center, Xi’an, 710061, Shaanxi, China

    Jianshui Zhang, Xinlin Chen & Yong Liu

  4. Department of Anatomy, Xi’an Medical University, Xi’an, 710021, Shaanxi, China

    Junfeng Zhang

  5. Department of Anesthesia of the Second Affiliated Hospital, Xi’an Jiaotong University Health Science Center, Xi’an, 710004, Shaanxi, China

    Pengbo Zhang

Authors
  1. Yingfang Tian
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  2. Yuanyuan Di
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  3. Jianshui Zhang
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  4. Xinlin Chen
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  10. Pengbo Zhang
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  11. Yong Liu
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Contributions

TY and LY conceived and designed the experiments. ZP and ZJ conducted pMCAO rat models. TY, DY, CX, and ZJ performed LCM experiments. FT, FJ, and SM did immunochemistry experiments. TY conducted qPCR experiments. TY wrote the article with the insightful advice from FA. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yingfang Tian or Yong Liu.

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The authors declare that they have no conflict of interest.

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Electronic Supplementary Material

Supplementary Figure 1

Representative coronal sections of rat brain stained with TTC at 24 h after pMCAO. Infarct brain tissue was in a white or pink color embedded in the healthy brain tissue of deep red color with TTC stain. The infarct size ranges from 30.3 to 38.6% in our study. (PNG 706 kb)

High Resolution Image (TIF 884 kb)

Supplementary Figure 2

The expression of Cd31/ Pecam1, NeuN/ Rbfox3, and Gfap mRNA in LCM-captured microvessels and neurons. M: Marker. (PNG 259 kb)

High Resolution Image (TIF 362 kb)

Supplementary Table 1

Genes that increased in microvessels at 2 h after ischemia (27 genes) (DOCX 18 kb)

Supplementary Table 2

Genes that increased in microvessels at 24 h after ischemia (23 genes) (DOCX 17 kb)

Supplementary Table 3

Genes that decreased in microvessels at 2 h after ischemia (8 genes) (DOCX 14 kb)

Supplementary Table 4

Genes that decreased in microvessels at 24 h after ischemia (4 genes) (DOCX 14 kb)

Supplementary Table 5

Genes that increased in neurons at 2 h after ischemia (16 genes) (DOCX 17 kb)

Supplementary Table 6

Genes that increased in neurons at 24 h after ischemia (26 genes) (DOCX 18 kb)

Supplementary Table 7

Genes that decreased in neurons at 2 h after ischemia (9 genes) (DOCX 17 kb)

Supplementary Table 8

Genes that decreased in neurons at 24 h after ischemia (5 genes) (DOCX 15 kb)

Supplementary Table 9

Genes that highly expressed in microvessels compared to neurons at control group (14 genes) (DOCX 20 kb)

Supplementary Table 10

Genes that highly expressed in neurons compared to microvessels at control group (9 genes) (DOCX 14 kb)

Supplementary Table 11

Genes that highly expressed in microvessels compared to neurons at 2 h after ischemia (17 genes) (DOCX 18 kb)

Supplementary Table 12

Genes that highly expressed in neurons compared to microvessels at 2 h after ischemia (10 genes) (DOCX 15 kb)

Supplementary Table 13

Genes that highly expressed in microvessels compared to neurons at 24 h after ischemia (16 genes) (DOCX 16 kb)

Supplementary Table 14

Genes that highly expressed in neurons compared to microvessels at 24 h after ischemia (10 genes) (DOCX 17 kb)

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Tian, Y., Di, Y., Zhang, J. et al. Angiogenic Gene Profiles in Laser-Microdissected Microvessels and Neurons from Ischemic Penumbra of Rat Brain. J Mol Neurosci 67, 643–653 (2019). https://doi.org/10.1007/s12031-019-01270-7

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  • DOI: https://doi.org/10.1007/s12031-019-01270-7

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