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Further study of CD31 protein and messenger ribonucleic acid expression in human cerebral vascular malformations
Ryunosuke Uranishi1, Nabil A Awadallah, Omolara O Ogunshola
1Neurovascular Surgery Program, Department of Neurosurgery, Yale University Medical School, New Haven, Connecticut, USA.
Objective:
In a previous study, we documented lower levels of immunoexpression of platelet endothelial cell (EC) adhesion molecule (CD31) in paraffin sections of cerebral cavernous malformations (CCMs), compared with arteriovenous malformations (AVMs) or normal brain tissue. We hypothesized that down-regulation of CD31 in CCMs might represent a distinctive phenotypic feature of ECs in this disease. To confirm this hypothesis, we further examined both protein and messenger ribonucleic acid (mRNA) expression of CD31, using immunohistochemical and in situ hybridization analyses, in fresh-frozen specimens of CCMs, AVMs, and control brain tissue.
Methods:
Fresh-frozen sections of four AVMs, five CCMs, and four control brain tissue specimens obtained from surgical resections were immunohistochemically stained with antibodies to von Willebrand factor and two distinct epitopes of CD31. In two AVMs, four CCMs, and three control brain tissue samples from the aforementioned group, the expression of CD31 mRNA was also examined by using in situ hybridization. Large (>100-microm) and small (<100-microm) vessels were counted and assessed for protein and mRNA expression.
Results:
In all tissues, ECs in the majority of vessels were immunopositive for CD31 with two distinct antibodies. CD31 mRNA was expressed in some but not all vessels in AVMs, CCMs, and control brain tissue. There were no statistically significant differences in CD31 protein or mRNA expression in CCMs, AVMs, and control brain tissue.
Conclusion:
The expression of CD31 in CCMs can be underestimated in paraffin sections. There does not seem to be a unique phenotypic differentiation of CD31 expression in ECs of CCMs or AVMs, compared with control brain tissue.
Insights
Platelet endothelial cell adhesion molecule (CD31) expression in cerebral cavernous malformations (CCMs) is not unique. This study found no significant differences in CD31 protein or mRNA levels in CCMs compared to other vascular malformations or normal brain tissue.
Area of Science:
- Neuroscience
- Vascular Biology
- Pathology
Background:
- Previous studies suggested lower CD31 immunoexpression in cerebral cavernous malformations (CCMs) paraffin sections.
- This observation raised the hypothesis of CD31 down-regulation as a distinctive feature of endothelial cells (ECs) in CCMs.
Purpose of the Study:
- To confirm the hypothesis of CD31 down-regulation in CCMs.
- To analyze CD31 protein and mRNA expression in fresh-frozen specimens of CCMs, arteriovenous malformations (AVMs), and control brain tissue.
Main Methods:
- Immunohistochemical staining for CD31 and von Willebrand factor on fresh-frozen tissue sections.
- In situ hybridization to assess CD31 mRNA expression.
- Quantification of CD31 expression in large and small vessels across CCMs, AVMs, and control tissues.
Main Results:
- CD31 protein and mRNA were expressed in endothelial cells across all examined tissues (CCMs, AVMs, controls).
- No statistically significant differences in CD31 protein or mRNA expression were found between CCMs, AVMs, and control brain tissue.
- CD31 expression in paraffin sections may underestimate its actual levels.
Conclusions:
- The expression of CD31 in CCMs is not uniquely down-regulated compared to AVMs or normal brain tissue.
- Underestimation of CD31 expression in paraffin-embedded tissues may have influenced previous findings.
- There is no distinct phenotypic differentiation of CD31 expression in the endothelial cells of CCMs or AVMs.
