Technical Reference #1656

1656.

Regulation of Synaptic Structure by Ubiquitin C-Terminal Hydrolase L1 Anna E. Cartier; Stevan N. Djakovic; Afshin Salehi; Scott M. Wilson; Eliezer Masliah; and Gentry N. Patrick, University of California, Neurobiology of Disease, 29(1656), (2009)
Link To Paper

Abstract:
Ubiquitin C-terminal hydrolase L1 (UCH-L1) is a deubiquitinating enzyme that is selectively and abundantly expressed in the brain and itsactivity is required for normal synaptic function. Here we show that UCH-L1 functions in maintaining normal synapt

Materials & Methods:
Primary neuronal cultures. Hippocampal neuron cultures were prepared from postnatal day 1 (P1) or P2 rat hippocampi as previously described (Patrick et al. 2003). Briefly for immunostaining experiments rat hippocampi were dissected dissociated by papain treatment and mechanical trituration and plated at medium density (45000 cells/cm 2) on poly-D-lysine-coated coverslips (12 mm in diameter) or glass-bottom dishes (MatTek; 35 mm). For biochemical experiments mixed hippocampal and cortical neurons were plated at high density on six-well plates (500000 cells per well) coated with poly-D-lysine. Cultures were maintained in B27 supplemented Neurobasal media (Invitrogen) until 14–21 d in vitro (DIV).

Microscopic Technique
Electron Microscopy

Cell Type(s)
BHK