Glutamate-induced Ca2+ influx in third-order neurons of salamander retina is regulated by the actin cytoskeleton.
Ligand-gated ion channels (ionotropic receptors) link to the cortical cytoskeleton via specialized scaffold proteins and thereby to appropriate signal transduction pathways in the cell. We studied the role of filamentous actin in the regulation of Ca influx through glutamate receptor-activated channels in third-order neurons of salamander retina. Staining by Alexa-Fluor 488-phalloidin, to visualize polymerized actin, we show localization of filamentous actin in neurites, and the membrane surrounding the cell soma. With Ca(2+) imaging we found that in dissociated neurons, depolymerization of filamentous actin by latrunculin A, or cytochalasin D significantly reduced glutamate-induced intracellular Ca(2+) accumulation to 53+/-7% of control value. Jasplakinolide, a stabilizer of filamentous actin, by itself slightly increased the glutamate-induced Ca(2+) signal and completely attenuated the inhibitory effect when applied in combination with actin depolymerizing agents. These results indicate that in salamander retinal neurons the actin cytoskeleton regulates Ca(2+) influx through ionotropic glutamate receptor-activated channels, suggesting regulatory roles for filamentous actin in a number of Ca(2+)-dependent physiological and pathological processes.
Akopian
A
A
Department of Ophthalmology, NYU School of Medicine, New York, NY 10016, USA. aa3@nyu.edu
Szikra
T
T
Cristofanilli
M
M
Krizaj
D
D
D000199:Actins / Q000378:metabolism / Q000502:physiology*; D000557:Ambystoma / Q000378:metabolism*; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology*; D003627:Data Interpretation, Statistical; D047630:Depsipeptides / Q000494:pharmacology; D018698:Glutamic Acid / Q000633:toxicity*; D066298:In Vitro Techniques; D018613:Microscopy, Confocal; D017470:Receptors, Glutamate / Q000187:drug effects / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000378:metabolism*; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D047630:Depsipeptides; D017470:Receptors, Glutamate; D013844:Thiazoles; D053778:Thiazolidines; C057531:jasplakinolide; D015638:Cytochalasin D; D018698:Glutamic Acid; C037067:latrunculin A; D002118:Calcium