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Expression of the neurofibromatosis type 2 gene in human tissues
M A den Bakker1, K J Vissers, A C Molijn
1Department of Pathology, Josephine Nefkens Institute, Rotterdam, The Netherlands.
Abstract:
The neurofibromatosis Type 2 tumor suppressor gene is implicated in the hereditary tumor syndrome NF2, hallmarked by bilateral vestibular schwannomas, meningiomas, and ocular non-neoplastic features. The gene product has characteristics of a membrane cytoskeleton-linking protein but the mechanism of tumor suppression by the NF2 protein remains to be elucidated. The NF2 gene is widely expressed in mouse and rat tissues. In humans, most of the expression data have accumulated through Northern blot analysis, RT-PCR and, more recently, Western blot analysis, providing information on whole tissues and organs rather than on specific cell types. We report here an extensive survey of NF2 gene expression in human tissues using a combination of mRNA in situ hybridization (mRNA ISH) and immunohistochemistry (IH) with a panel of monoclonal antibodies (MAbs) supplemented by tissue immunoprecipitation experiments with affinity-purified polyclonal antibodies. Expression was observed in many different cell types, most of which appear functionally normal in individuals affected by NF2. Surprisingly, expression could not be consistently documented in Schwann cells and arachnoidal cells by IH or by mRNA ISH in formalin-fixed tissue. However, consistent immunostaining of Schwann cells was seen in frozen sections. (J Histochem Cytochem 47:1471-1479, 1999)
Insights
The neurofibromatosis type 2 (NF2) gene product is crucial for tumor suppression. This study extensively surveyed NF2 gene expression in human tissues, revealing widespread cellular distribution.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Neurofibromatosis type 2 (NF2) is a hereditary tumor syndrome linked to the NF2 tumor suppressor gene.
- The NF2 protein functions as a membrane cytoskeleton-linking protein, but its tumor suppression mechanism is unclear.
- Previous human NF2 gene expression data relied on whole-tissue analyses.
Purpose of the Study:
- To extensively survey NF2 gene expression in various human cell types.
- To investigate NF2 protein localization and expression patterns.
- To identify cell-specific expression of the NF2 gene.
Main Methods:
- Utilized mRNA in situ hybridization (mRNA ISH) and immunohistochemistry (IH) on human tissues.
- Employed a panel of monoclonal antibodies (MAbs) and polyclonal antibodies for immunoprecipitation.
- Examined both formalin-fixed and frozen tissue sections.
Main Results:
- NF2 gene expression was observed in numerous human cell types, many appearing functionally normal in NF2 patients.
- Immunohistochemistry and mRNA ISH on formalin-fixed tissues did not consistently detect NF2 expression in Schwann cells and arachnoidal cells.
- Consistent NF2 immunostaining was observed in Schwann cells using frozen tissue sections.
Conclusions:
- The NF2 gene is broadly expressed across diverse human cell types.
- Discrepancies in Schwann cell NF2 detection highlight the importance of tissue fixation methods.
- Further research is needed to fully elucidate the tumor suppression mechanisms of the NF2 protein.