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Journal of Molecular Neuroscience

, Volume 67, Issue 4, pp 643–653 | Cite as

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

  • Yingfang TianEmail author
  • Yuanyuan Di
  • Jianshui Zhang
  • Xinlin Chen
  • Ting Feng
  • Frank Adu-Nti
  • Meimei Shi
  • Juan Fan
  • Junfeng Zhang
  • Pengbo Zhang
  • Yong LiuEmail author
Article
  • 78 Downloads

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.

Keywords

Cerebral ischemia Laser capture microdissection Angiogenesis Microvessel Neuron 

Notes

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.

Authors’ 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.

Funding Information

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.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12031_2019_1270_Fig7_ESM.png (706 kb)
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)

12031_2019_1270_MOESM1_ESM.tif (885 kb)
High Resolution Image (TIF 884 kb)
12031_2019_1270_Fig8_ESM.png (260 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)

12031_2019_1270_MOESM2_ESM.tif (363 kb)
High Resolution Image (TIF 362 kb)
12031_2019_1270_MOESM3_ESM.docx (18 kb)
Supplementary Table 1 Genes that increased in microvessels at 2 h after ischemia (27 genes) (DOCX 18 kb)
12031_2019_1270_MOESM4_ESM.docx (18 kb)
Supplementary Table 2 Genes that increased in microvessels at 24 h after ischemia (23 genes) (DOCX 17 kb)
12031_2019_1270_MOESM5_ESM.docx (15 kb)
Supplementary Table 3 Genes that decreased in microvessels at 2 h after ischemia (8 genes) (DOCX 14 kb)
12031_2019_1270_MOESM6_ESM.docx (14 kb)
Supplementary Table 4 Genes that decreased in microvessels at 24 h after ischemia (4 genes) (DOCX 14 kb)
12031_2019_1270_MOESM7_ESM.docx (18 kb)
Supplementary Table 5 Genes that increased in neurons at 2 h after ischemia (16 genes) (DOCX 17 kb)
12031_2019_1270_MOESM8_ESM.docx (19 kb)
Supplementary Table 6 Genes that increased in neurons at 24 h after ischemia (26 genes) (DOCX 18 kb)
12031_2019_1270_MOESM9_ESM.docx (17 kb)
Supplementary Table 7 Genes that decreased in neurons at 2 h after ischemia (9 genes) (DOCX 17 kb)
12031_2019_1270_MOESM10_ESM.docx (15 kb)
Supplementary Table 8 Genes that decreased in neurons at 24 h after ischemia (5 genes) (DOCX 15 kb)
12031_2019_1270_MOESM11_ESM.docx (21 kb)
Supplementary Table 9 Genes that highly expressed in microvessels compared to neurons at control group (14 genes) (DOCX 20 kb)
12031_2019_1270_MOESM12_ESM.docx (15 kb)
Supplementary Table 10 Genes that highly expressed in neurons compared to microvessels at control group (9 genes) (DOCX 14 kb)
12031_2019_1270_MOESM13_ESM.docx (19 kb)
Supplementary Table 11 Genes that highly expressed in microvessels compared to neurons at 2 h after ischemia (17 genes) (DOCX 18 kb)
12031_2019_1270_MOESM14_ESM.docx (16 kb)
Supplementary Table 12 Genes that highly expressed in neurons compared to microvessels at 2 h after ischemia (10 genes) (DOCX 15 kb)
12031_2019_1270_MOESM15_ESM.docx (16 kb)
Supplementary Table 13 Genes that highly expressed in microvessels compared to neurons at 24 h after ischemia (16 genes) (DOCX 16 kb)
12031_2019_1270_MOESM16_ESM.docx (18 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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Key Laboratory of Modern Teaching TechnologyMinistry of EducationXi’anChina
  2. 2.College of Life SciencesShaanxi Normal UniversityXi’anChina
  3. 3.Institute of Neurobiology, School of Basic Medical SciencesXi’an Jiaotong University Health Science CenterXi’anChina
  4. 4.Department of AnatomyXi’an Medical UniversityXi’anChina
  5. 5.Department of Anesthesia of the Second Affiliated HospitalXi’an Jiaotong University Health Science CenterXi’anChina

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