title: Imaging of calcineurin activated by long-term depression-inducing synaptic inputs in living neurons of rat visual cortex.
abstract: Long-term depression (LTD) of synaptic transmission is induced by low-frequency stimulation (LFS) of afferents lasting for a long time, typically for 10-15 min, in neocortical and hippocampal slices. It is suggested that calcineurin, Ca2+/calmodulin-dependent protein phosphatase, plays a role in the induction of LTD, based on the results that pharmacological or genetic manipulation of calcineurin activity interfered in its induction. However, questions as to why it takes so long to induce LTD and in which compartment of neurons calcineurin is activated remain unanswered. With a fluorescent indicator for calcineurin activity, we visualized the spatiotemporal pattern of its activation in living neurons in layer II/III of visual cortical slices of rats during the LFS of layer IV that induced LTD of synaptic responses. During LFS, the fluorescence intensity gradually increased with a latency of a few minutes in dendrites and soma of neurons, and remained increased during the whole observation period (10-25 min) after LFS. The onset latency of the increase in the soma was slower than that in the distal dendritic region. The LFS-induced rise in fluorescence was not observed in neurons which were loaded with inhibitors of calcineurin, indicating that the intensity of fluorescence reflects calcineurin activity. Control stimulation at 0.05 Hz and theta-burst stimulation did not significantly change the intensity of fluorescence. Only LFS-type inputs effectively activate calcineurin in postsynaptic neurons in an augmenting manner, and such a time-consuming activation of calcineurin may be a reason why long-lasting LFS is necessary for the induction of LTD.
authors.affiliation: Division of Neurophysiology, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita 565-0871, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yasuda
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Higashi
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Kudo
authors.forename: Takafumi
authors.initials: T
authors.lastname: Inoue
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Hata
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Mikoshiba
authors.forename: Tadaharu
authors.initials: T
authors.lastname: Tsumoto
mesh_terms: D000818:Animals; D019703:Calcineurin / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004558:Electric Stimulation; D004594:Electrophysiology; D004789:Enzyme Activation / Q000502:physiology*; D019706:Excitatory Postsynaptic Potentials / Q000502:physiology; D007202:Indicators and Reagents; D017774:Long-Term Potentiation / Q000187:drug effects / Q000502:physiology*; D009473:Neuronal Plasticity / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D009924:Organ Culture Techniques; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014793:Visual Cortex / Q000187:drug effects / Q000502:physiology*
chemical_list: D007202:Indicators and Reagents; D019703:Calcineurin