Technical Reference #1756

1756.

Rapid Assembly of Functional Presynaptic Boutos Triggered by Adhesive Contacts Anna Lisa Lucido; Fernando Suarez Sanchez; Peter Thostrup; Adam V. Kwiatkowski; Sergio Leal-Ortiz; Gopakumar Gopalakrishnan; Dalinda Liazoghli; Wiam Belkaid; R. Bruce Lennox; Peter Grutter; Craig C. Garner; David R. Colman, McGill University, Journal of Neuroscience, 29(1756), (2009)
Link To Paper

Materials & Methods:
Neuronal culture. All animal experimentation was approved by the institutional animal care committee and conformed to the guidelines of the Canadian Council of Animal Care. All culture media was purchased from Invitrogen. Hippocampi were dissected from embryonic day 17–18 (E17–E18) Sprague Dawley rat embryos (Charles River) as described previously (Benson and Tanaka 1998). For immunocytochemistry cells were plated at a density of 2.0 –2.5 104 cm 1 on PDL (Sigma-Aldrich)- coated glass coverslips. For electron microscopy cells were plated on PDL-treated four-well plates (Nunc VWR) at a density of 5.0 104 cells per well. All cells were cultured in serum-free Neurobasal medium supplemented with L-glutamine and B-27. Transfection of plasmidDNAwas performed using Lipofectamine (Invitrogen) according to the company protocol and transfected cells were incubated for a minimum of 48 h before experimentation. Mouse synaptophysin-EGFP plasmid was a gift from Dr. Edward Ruthazer (McGill University) and pEGFP-C1 plasmid was purchased from Clontech.

Microscopic Technique
fluorescence; atomic force microscopy; confocal microscopy

Cell Type(s)
Hippocampal cultures