Kalirin, a Key Player in Synapse Formation, Is Implicated in Human Diseases
Figure 3
Expression of Kalirin-7 shRNA causes a reduction in spine density and spine size in cultured hippocampal neurons. Cultured hippocampal neurons prepared at E20 were transfected with vector encoding a membrane-tethered version of GFP (pmGFP) alone [(a), spine density 8.9 ± 0.8/10 μm], pmGFP plus a scrambled shRNA [(b), spine density 9.2 ± 0.9/10 μm], or pmGFP plus pSIREN-Kalirin-7 shRNA [(c), spine density 5.2 ± 0.6/10 μm] on the day of culture preparation. The specificity of pSIREN-Kalirin-7 shRNA was determined previously [19]. DsRed marks transfected neurons expressing the shRNA. Images were collected on day 20 with an LSM510 confocal microscope. Expression of Kalirin-7 shRNA, not scrambled shRNA, caused a 80% decrease in Kalirin-7 staining (not shown). Spine density was determined using Metamorph (Molecular Devices, Downington, PA) as described [54]. Scale bar = 5 μm.
