Figure 3.
dHIP14YFP localizes to synapses in optic lobe, central brain, and larval ventral nerve cord neurons. A–F, Cryostat sections of the optic lobes of a control fly (No dHIP14YFP) (A–C) and a dHIP14YFP fly (D–F) triple-labeled with the synaptic vesicle marker neuronal-synaptobrevin (N-syb) (A, D), a Golgi marker (B, E), and anti-YFP (C, F). dHIP14YFP distributes to the synaptic neuropil of the optic lobe in a pattern very similar to the synaptic vesicle marker N-syb and clearly distinct from the Golgi marker. Background levels of fluorescence for the anti-YFP antibody can be observed in C. lam, Lamina; med, medula; lob, lobula; lob pl, lobula plate. Scale bar, 50 μm. G–L, High-resolution images of cryostat sections of the cell body (G–I) and lamina (J–L) regions of photoreceptors of a dHIP14YFP fly double-immunolabeled with a Golgi marker (G, J), anti-YFP (H, K), and the overlay (I, L). The arrow (G–I) points to the same cell body in each. dHIP14YFP is not detected in either the photoreceptor or presumptive lamina cell body Golgi, but is present in non-neuronal cell bodies adjacent to the lamina. Scale bar, 20 μm. M–O, Cryostat sections of the central brain of a dHIP14YFP fly triple-labeled for the synaptic vesicle marker N-syb (M), a Golgi marker (N), and anti-YFP (O). Similar to the optic lobe, the vast majority of dHIP14YFP localizes in the pattern of the synaptic vesicle marker, but note the overlap with the Golgi marker in adjacent non-neuronal regions. Scale bar, 50 μm. P–R, First instar ventral nerve cord of a dHIP14YFP larvae (VNC) labeled with Synapsin (P), direct dHIP14YFP fluorescence (Q), and the overlay (R). The majority of dHIP14YFP colocalizes with the synaptic vesicle marker Synapsin in the synaptic neuropil region of the VNC. S–U, First instar VNC of a dHIP14YFP larvae labeled with a Golgi marker (S), direct dHIP14YFP fluorescence (T), and the overlay (U). The Golgi marker is restricted to the cell body region of the VNC, whereas a portion of dHIP14YFP is present in the cell body, but most distributes to the synaptic neuropil. Scale bar, 50 μm. Genotypes are as follows: yw; cn bw sp/cn bw sp (A–C), yw; cn bw sp/cn bw sp; dHIP14ex11, dHIP14YFP/dHIP14ex11, dHIP14YFP (D–O), yw; dHIP14ex11, dHIP14YFP/dHIP14ex11, dHIP14YFP (P–U). lam, Lamina; med, medulla; lob, lobula; lob pl, lobula plate.