Fig. 7.
Colocalization of recombinant Mint1 and CASK with N-type channels in mature hippocampal neurons. A–D, High-density hippocampal neuronal cultures transfected with GFP-Mint1 (A, B) and GFP-CASK (B, D) plasmids at 9 DIV. At 3–5 d after transfection, the localization of GFP-Mint1 or GFP-CASK (green) and endogenous MAP2 (red) was determined. The somatodendritic domain (A, C) and distal axonal segment (B, D) of transfected cells are shown. Proximal axonal segments are indicated by arrows (A, C). Presynaptic clusters of GFP-Mint1 (B) and GFP-CASK (D) associated with axons of transfected neurons apposed to dendrites (MAP2-positive) are indicated by small arrows. Scale bars, 20 μm. E, F, High-density mature hippocampal neurons cotransfected with HA-α1B/α2δ/β3 and GFP-Mint1 (E) or GFP-CASK (F) at 9 DIV. At 4 d after transfection, the axonal distribution of HA-α1B (top panel,red in merged image) and GFP-Mint1 (E, center panel, green inmerged image) or GFP-CASK (F,center panel, green in merged image) was determined. Synaptic clusters of HA-α1B, GFP-Mint1, and GFP-CASK are indicated bysmall arrows. Axonal growth cones are indicated byasterisks in E. Scale bars, 20 μm.