Fig. 1. An NCS-1-related protein is expressed in L. stagnalis neurons. (A) Protein sequence alignment of the full NCS-1 clone from L. stagnalis (DQ099793) and the homologous proteins from A. californica (Q16981), Rattus norvegicus (P62168), Drosophila melanogaster (NP_996502) and Caenorhabditis elegans (P36608). Alignment was performed in ClustalW and the plot made in GeneDoc. (B) Western blot analysis of whole L. stagnalis and rat brains. Polyclonal anti-rat NCS-1 antibody identified a single band migrating at 23 kDa in both rat and L. stagnalis ganglia preparations. (C,D) Confocal imaging of anti-NCS-1 reactivity in a L. stagnalis PeA neuron cultured for 3 days in CM. Epifluorescent image of PeA cell stained with the anti-rat NCS-1 antibody show immunoreactivity throughout the neuron (C). Analysis of cellular distribution of NCS-1 reactivity in the different regions of the cell (D, n=5 cells). The data are presented as mean±s.e.m. Asterisk indicates a significant difference (P<0.05) compared with that in the primary neurites. Relativity intensities compared to primary neurites are 2.95±0.39 for soma, 1.91±0.31 for branch points, 1.27±0.29 for secondary neurites and 1.85±0.10 for growth cones.