Degradation of Filamin Induces Contraction of Vascular Smooth Muscle Cells in Type-I Collagen Matrix Honeycombs
| Autor: | Itsuko Ishii, Masashi Uchida, Fumiko Yamamoto, Yuji Nakayama, Noritaka Ariyoshi, Kaori Tashiro, Takayoshi Suzuki, Mitsukazu Kitada, Kaori Hirata |
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| Rok vydání: | 2011 |
| Předmět: |
Male
Vascular smooth muscle Physiology Filamins macromolecular substances Filamin Collagen Type I Muscle Smooth Vascular Mice Contractile Proteins Western blot medicine Animals Cytoskeleton Actin DNA Primers Base Sequence biology medicine.diagnostic_test Reverse Transcriptase Polymerase Chain Reaction Microfilament Proteins Calpain respiratory system musculoskeletal system respiratory tract diseases Cell biology Cytoplasm cardiovascular system biology.protein Rabbits Type I collagen |
| Zdroj: | Cellular Physiology and Biochemistry. 27:669-680 |
| ISSN: | 1421-9778 1015-8987 |
| DOI: | 10.1159/000330076 |
| Popis: | Background: Dedifferentiated rabbit vascular smooth muscle cells (SMCs) exhibit similar features to differentiated SMCs when cultured in three-dimensional matrices of type-I collagen called “honeycombs,” but the mechanism is unknown. The role of filamin, an actin-binding protein that links actin filaments in SMCs, was investigated. Methods: Filamin and other related proteins were detected by western blot analysis and immunofluorescence staining. Honeycomb size was measured to confirm the contraction of SMCs. Results: Full-length filamin was expressed in subconfluent SMCs cultured on plates; however, degradation of filamin, which might be regulated by calpain, was observed in confluent SMCs cultured on plates and in honeycombs. While filamin was co-localized with β-actin in subconfluent SMCs grown on plates, filamin was detected in the cytoplasm in SMCs cultured in honeycombs, and degraded filamin was mainly detected in the cytoplasmic fraction of these cells. In addition, β-actin expression was low in the cytoskeletal fraction of SMCs cultured in honeycombs compared with cells cultured on plates, and the size of the honeycombs used for culturing SMCs was significantly reduced. Conclusion: These data suggest that degradation of filamin in SMCs cultured in honeycombs induces structural weakness of β-non-muscle actin filaments, thereby permitting SMCs in honeycombs to achieve contractility. |
| Databáze: | OpenAIRE |
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