Capturing in vivo dynamics of the actin cytoskeleton stimulated by auxin or light.
We present here a transient expression system that allows the response of actin microfilaments to physiological stimuli (changes in auxin content, light) to be observed in single cells in vivo. Etiolated, intact rice seedlings are attached to glass slides, transfected biolistically with talin fused to yellow-fluorescent protein to visualize actin microfilaments, and either treated with auxin or irradiated. The talin marker labels distinct populations of actin that are differentially expressed depending on the physiological state of the coleoptile (active elongation versus ceased elongation). Whereas longitudinal transvacuolar bundles prevail in cells that have ceased to elongate, fine cortical strands are characteristic for elongating cells. The visualized actin structures remain dynamic and responsive to signals. Exogenous auxin triggers a loosening of the bundles and an extension of the cortical strands, whereas irradiation reorientates cortical strands into longitudinal arrays. These responses correspond in quality and timing to the signal responses inferred previously from fixed specimens and biochemical studies. In big advantage over those methods it is now possible to observe them directly at the single cell level. Thus, the rice coleoptile system can be used as a convenient model to study actin dynamics in vivo, in response to physiologically relevant stimuli.
Holweg
Carola
C
Institut für Biologie II, Schänzlestrasse 1, D-79104 Freiburg, Germany. carola.holweg@biologie.uni-freiburg.de
Süsslin
Christina
C
Nick
Peter
P
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D000199:Actins / Q000378:metabolism*; D001426:Bacterial Proteins; D001681:Biological Assay / Q000379:methods; D018548:Cotyledon / Q000187:drug effects / Q000254:growth & development / Q000528:radiation effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D007210:Indoleacetic Acids / Q000378:metabolism* / Q000494:pharmacology; D008027:Light; D008164:Luminescent Proteins; D008954:Models, Biological; D012275:Oryza / Q000187:drug effects / Q000254:growth & development* / Q000528:radiation effects; D036226:Seedlings / Q000187:drug effects / Q000254:growth & development* / Q000528:radiation effects
D000199:Actins; D001426:Bacterial Proteins; D007210:Indoleacetic Acids; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria