Eph receptor tyrosine kinases regulate astrocyte cytoskeletal rearrangement and focal adhesion formation.
EphA4 null mice have impaired astrocytic gliosis following spinal cord injury. This may be because of altered cytoskeletal regulation and is examined herein using cultured astrocytes from wildtype and EphA4 null mice. Under basal conditions EphA4 null astrocytes appeared relatively normal but following stimuli resulting in cytoskeletal rearrangement, EphA4 null cells responded more slowly. When F-actin stress fibers were collapsed using the Rho kinase inhibitor HA1077, fewer EphA4 null cells showed stress fiber collapse in response to HA1077 and recovered stress fibers more slowly following HA1077 removal. EphA4 null astrocytes were less adherent and had smaller focal adhesions, while activation of Eph receptors with ephrin-A5-Fc increased the numbers of focal adhesions in both wildtype and knockout astrocytes following serum starvation. Using scratch wound assays, EphA4 null astrocytes invading the scratch showed impaired glial fibrillary acidic protein expression, particularly in proliferative cells. Astrocytes did not express Ephexin, a major Eph-interacting Rho guanine exchange factor, but they expressed Vav proteins, with lower levels of phospho-Vav in EphA4 null compared to wildtype astrocytes. This may contribute to the slower cytoskeletal responses generally observed in the EphA4 null astrocytes. Eph receptor signaling therefore regulates astrocyte reactivity through modulation of cytoskeletal responses.
Puschmann
Till B
TB
Centre for Neuroscience, The University of Melbourne, Victoria, Australia.
Turnley
Ann M
AM
D000199:Actins / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism* / Q000648:ultrastructure; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D004791:Enzyme Inhibitors / Q000494:pharmacology; D036386:Ephrin-A5 / Q000494:pharmacology; D022001:Focal Adhesions / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005911:Gliosis / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D051567:Proto-Oncogene Proteins c-vav / Q000378:metabolism; D036122:Receptor, EphA4 / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D013119:Spinal Cord Injuries / Q000378:metabolism / Q000503:physiopathology; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
D000199:Actins; D004791:Enzyme Inhibitors; D036386:Ephrin-A5; D005904:Glial Fibrillary Acidic Protein; D051567:Proto-Oncogene Proteins c-vav; D036122:Receptor, EphA4; D054460:rho-Associated Kinases