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Laser Capture Microdissection of Enriched Populations of Neurons or Single Neurons for Gene Expression Analysis After Traumatic Brain Injury
Published on: April 10, 2013
[A primary study on the gene expression profiling of human brain contusion by cDNA microarray]
Zhou Yi-wu1, Zhang Yi-gu, Liu Yan
1Faculty of Forensic Medicine, Tongji Medical College, Huazfong University of Science and Technology, Wuhan 430030, China. yiwu@public.wh.hb.cn
Objective:
To screen the differential expression of oncogenes and tumors suppressed genes(OTS genes) after human brain contusion by cDNA microrarray.
Methods:
The total RNAs isolated from normal and contusion human brain tissues were purified by Oligotex to obtain mRNAs. Both sources of mRNAs were reversely transcribed to cDNAs with the incorporation of fluorescent dUTP to prepare the hybridization probes. The probe from normal tissue and the contusion brain tissue were labeled with Cy3-dUTP and Cy5-dUTP respectively. The mixed probes were hybridized to the BioDoor Chip OTS-2.2S, a cDNA microarray which contains 227 oncogenes and tumors suppressed genes. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed differences between two tissues.
Results:
Among the 227 target genes, 3 genes including Human carcinoma associated HOJ-1 (HoJ-1), Human KIAAOO65 gene,Human retinoblastoma related protein (p107) gene, showed distinct deference in expression level between the human brain contusion tissue and normal tissue.
Conclusion:
The 3 genes in the brain contusion was significantly the differential expression by OTS 2.2S cDNA microarray. Further analysis of these genes will be helpful to understand the molecular mechanism of brain injury and utilization in forensic medicine.
Insights
Differential gene expression in human brain contusion was screened using cDNA microarray. Three genes, including HOJ-1, KIAA0065, and p107, showed significant expression changes, offering insights into brain injury mechanisms.
Area of Science:
- Molecular Biology
- Genomics
- Neuroscience
Context:
- Human brain contusion involves complex molecular changes.
- Identifying differentially expressed genes is crucial for understanding injury.
- Oncogenes and tumor suppressor genes (OTS genes) play roles in cellular processes.
Purpose:
- To screen for differential expression of OTS genes in human brain contusion tissue.
- To identify specific genes altered in expression following brain injury.
- To utilize cDNA microarray technology for comprehensive gene expression analysis.
Summary:
- A cDNA microarray (BioDoor Chip OTS-2.2S) containing 227 OTS genes was employed.
- Total RNAs from normal and contusion human brain tissues were analyzed.
- Three genes—HOJ-1, KIAA0065, and p107—exhibited significant differential expression.
Impact:
- The identified genes provide potential biomarkers for brain injury.
- Understanding these molecular alterations can elucidate the pathogenesis of brain trauma.
- Findings may contribute to forensic medicine applications in diagnosing brain injuries.
