Figure 6. ATG-9 Transport to AIY PresynapticZones Requires KIF1A/UNC-104(A) Schematic of the nematode nerve ring (green, encircled by a dotted line) in C. elegans.(B) Schematic of the
atg-9 genomic region in a wild-type genome (top) and in the enhanced GFP CRISPR knockin (bottom). (C and D) Location of the nerve ring, referenced with a transmitted light image (C) and visualized with GFP::RAB-3 (pseudocolored red) expressed panneuronally with Prab-3 in a wild-type animal (D).(E) Visualization of the subcellular localization of panneuronal ATG-9 (ATG-9::GFP rescuing array expressed with Punc-14). (F-K) Distribution of endogenous ATG-9 as visualized by CRISPR insertion of enhanced GFP at the C terminus of the genomic
atg-9 (schematized in (B)) in adult (F-H) and embryo (I-K) in wild-type (G and J) and
unc-104(
e1265) mutant (H and K) animals. Anterior end of the embryo (head) is indicated with an asterisk in (I)-(K).(L-Q) Distribution of synaptic vesicles (visualized with mCh::RAB-3) (L and O) and ATG-9::GFP (M and P) in AIY (merge in N and Q) in wild-type(L-N) and
unc-104(
e1265) mutant animals (O-Q). In (L)-(Q), the asterisk denotes the location of the AIY cell body; the dashed box encloses AIYzone 2. Each image is a maximal projection of a confocal z stack. Scale bar in (D) for (C)-(H), in (J) for (I)-(K), and in (L) for (L)-(Q), 5 um.