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Characterization of Diaphanous-related formin FMNL2 in human tissues
Maria Gardberg1, Kati Talvinen, Katja Kaipio
1Department of Pathology, University of Turku and Turku University Central Hospital, Turku, Finland. maria.gardberg@utu.fi
Background:
Diaphanous-related formins govern actin-based processes involved in many cellular functions, such as cell movement and invasion. Possible connections to developmental processes and cellular changes associated with malignant phenotype make them interesting study targets. In spite of this, very little is known of the tissue distribution and cellular location of any mammalian formin. Here we have carried out a comprehensive analysis of the formin family member formin -like 2 (FMNL2) in human tissues.
Results:
An FMNL2 antibody was raised and characterized. The affinity-purified FMNL2 antibody was validated by Western blotting, Northern blotting, a peptide competition assay and siRNA experiments. Bioinformatics-based mRNA profiling indicated that FMNL2 is widely expressed in human tissues. The highest mRNA levels were seen in central and peripheral nervous systems. Immunohistochemical analysis of 26 different human tissues showed that FMNL2 is widely expressed, in agreement with the mRNA profile. The widest expression was detected in the central nervous system, since both neurons and glial cells expressed FMNL2. Strong expression was also seen in many epithelia. However, the expression in different cell types was not ubiquitous. Many mesenchymal cell types showed weak immunoreactivity and cells lacking expression were seen in many tissues. The subcellular location of FMNL2 was cytoplasmic, and in some tissues a strong perinuclear dot was detected. In cultured cells FMNL2 showed mostly a cytoplasmic localization with perinuclear accumulation consistent with the Golgi apparatus. Furthermore, FMNL2 co-localized with F-actin to the tips of cellular protrusions in WM164 human melanoma cells. This finding is in line with FMNL2's proposed function in the formation of actin filaments in cellular protrusions, during amoeboid cellular migration.
Conclusion:
FMNL2 is expressed in multiple human tissues, not only in the central nervous system. The expression is especially strong in gastrointestinal and mammary epithelia, lymphatic tissues, placenta, and in the reproductive tract. In cultured melanoma cells, FMNL2 co-localizes with F-actin dots at the tips of cellular protrusions.
Insights
Formin-like 2 (FMNL2) is widely expressed across human tissues, particularly in the nervous system and epithelia. This protein localizes to the cytoplasm and actin-rich cellular protrusions, suggesting a role in cell migration.
Area of Science:
- Cell Biology
- Molecular Biology
- Human Physiology
Background:
- Diaphanous-related formins regulate essential cellular functions like movement and invasion.
- Understanding mammalian formin tissue distribution and cellular localization is limited.
- Formin-like 2 (FMNL2) is a key formin family member with potential roles in development and malignancy.
Purpose of the Study:
- To comprehensively analyze the tissue distribution and cellular localization of formin-like 2 (FMNL2) in human tissues.
- To validate an antibody for FMNL2 detection.
- To investigate the subcellular localization of FMNL2 and its relationship with F-actin.
Main Methods:
- Antibody generation and characterization (Western blotting, Northern blotting, peptide competition, siRNA).
- Bioinformatics-based mRNA profiling for FMNL2 expression.
- Immunohistochemical analysis of FMNL2 in 26 human tissues.
- Subcellular localization studies in cultured cells and melanoma cell lines.
Main Results:
- FMNL2 mRNA is widely expressed, with highest levels in the central and peripheral nervous systems.
- Immunohistochemistry confirmed widespread FMNL2 expression, notably in neurons, glial cells, and various epithelia.
- FMNL2 exhibits cytoplasmic localization, with perinuclear accumulation and co-localization with F-actin at cellular protrusion tips in melanoma cells.
Conclusions:
- FMNL2 is expressed in diverse human tissues beyond the central nervous system, including strong presence in gastrointestinal and mammary epithelia, lymphatic tissues, placenta, and reproductive tract.
- FMNL2 localizes to the cytoplasm and actin-rich cellular protrusions, supporting its role in actin filament formation and cell migration.
- The study provides crucial insights into the expression pattern and subcellular localization of FMNL2 in human tissues.
