Biphasic and directed translocation of protein kinase Cα inside cultured endothelial cells before migration
| Autor: | Tomoya Shimada, Susumu Kudo, Chihiro Kora, Toshihiro Sera, Masataka Arai, Kazuhiro Nakashima |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Cell Biophysics Biology Protein kinase Cα Biochemistry Cell membrane lcsh:Biochemistry 03 medical and health sciences 0302 clinical medicine Endothelial cell PKCα protein kinase Cα SA stretch-activated Fluorescence microscope medicine lcsh:QD415-436 Protein kinase A lcsh:QH301-705.5 Mechanical stimulation integumentary system EC endothelial cell Mechanical injury Cell migration Fusion protein Cell biology Endothelial stem cell Cytosol 030104 developmental biology medicine.anatomical_structure lcsh:Biology (General) Fura-2–AM Fura-2–acetoxymethyl ester 030217 neurology & neurosurgery Research Article |
| Zdroj: | Biochemistry and Biophysics Reports, Vol 12, Iss C, Pp 91-97 (2017) Biochemistry and Biophysics Reports |
| ISSN: | 2405-5808 |
| Popis: | Mechanical wounding of an endothelial monolayer induces an immediate Ca2+ wave. Several hours later, the denuded area is covered by endothelial cells (ECs) that migrate to the wound. This migration process is closely related to protein kinase Cα (PKCα), a Ca2+-dependent protein that translocates from the cytosol to the cell membrane. Because the cells adjacent to the wounded area are the first to migrate into the wound, we investigated whether a mechanical wound immediately induces PKCα translocation in adjacent cells. We monitored Ca2+ dynamics and PKCα translocation simultaneously using fluorescent microscopy. For this simultaneous observation, we used Fura-2–acetoxymethyl ester to visualize Ca2+ and constructed a green fluorescent protein-tagged fusion protein to visualize PKCα. Mechanical wounding of the endothelial monolayer induced an immediate Ca2+ wave in cells adjacent to the wounded cells before their migration. Almost concurrently, PKCα in the neighboring cells translocated to the cell membrane, then accumulated at the periphery near the wounded cell. This report is the first description of this biphasic and directed translocation of PKCα in cells before cell migration. Our results may provide new insights into the directed migration of ECs. Highlights • We wounded a single endothelial cell (EC) and investigated the distribution of protein kinase Cα (PKCα) in adjacent ECs. • Initially, PKCα translocates to the cell membrane. • Thereafter, PKCα accumulates at the cell periphery adjacent to the wounded cell. |
| Databáze: | OpenAIRE |
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