Under copyright Constraint(s) on Use: This work is protected by the U.S. Copyright Law (Title 17, U.S.C.). Use of this work beyond that allowed by "fair use" or any license applied to this work requires written permission of the copyright holder(s). Responsibility for obtaining permissions and any use and distribution of this work rests exclusively with the user and not the UC San Diego Library. Inquiries can be made to the UC San Diego Library program having custody of the work. Use: This work is available from the UC San Diego Library. This digital copy of the work is intended to support research, teaching, and private study.
Rights Holder and Contact
UC Regents
Description
Rat hippocampal neurons grown in vitro, fixed after 23 days in culture, and immunostained for MAP2 (red) to reveal the dendritic arbor, and localization of α Calcium Calmodulin Kinase II (green), a multifunctional kinase that has high concentrations at excitatory postsynaptic sites and presynaptic terminals, and is involved in regulating synaptic plasticity. Detailed methods: Embryonic rat hippocampal neurons were prepared as previously described (see Kaech and Banker, 2006, Nat Protoc). Cells were prepared for fluorescent staining as previously described (Withers and Banker, 1998, in Culturing Nerve Cells, MIT Press). Briefly, cells were fixed (4% formaldehyde, 4% sucrose in phosphate buffered saline, pH 7.4, warmed to 37°C prior to fixation, 15 minutes), permeabilized with 0.25% Triton (7 minutes) and immunostained for alpha-CamKII (clone 6G9, Abcam, with Alexa 488 conjugated secondary, excitation, 494, emission, 519 [Invitrogen, Molecular Probes]) and MAP2 (Ab266 from S. Halpain, with DyLight549 conjugated secondary, excitation, 555, emission, 568, [Jackson Immunoresearch]). Images were acquired with a Leica DMRA microscope with a mercury arc lamp, a 40X lens (HCX PL Fluotar, NA 0.75), Leica GFP filter set (excitation, BP 470/40; dichromatic mirror, 500; suppression filter, BP 525/50); Leica N3 filter set (excitation, BP546/12; dichromatic mirror, 565, suppression filter, BP 600/40), Photometrics CoolSnap ES CCD camera and MetaMorph software. This merged image was generated with the MetaMorph color combine function. Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Brandner, Dieter; Withers, Ginger (2021). CIL:8733, Rattus, multipolar neuron. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0CC10P0
Type
image
Identifier
ark:/20775/bb1031441w
Language
No linguistic content
Subject
Dendrite development Calmodulin-dependent protein kinase activity Establishment or maintenance of cell polarity Cellular developmental process Synapse Microtubule cytoskeleton Dendrite Multipolar neuron Axon Rattus Cell Image Library Group ID: 7783
If you're wondering about permissions and what you can do with this item, a good starting point is the "rights information" on this page. See our terms of use for more tips.
Share your story
Has Calisphere helped you advance your research, complete a project, or find something meaningful? We'd love to hear about it; please send us a message.
