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. 2018 Oct 31;210(3):947–968. doi: 10.1534/genetics.118.301466

Figure 9.

Figure 9

Sentryn shows a SYD-2-dependent enrichment at active zones. [see above]. (A) Rectangle in drawing shows region imaged. Representative images showing colocalization of Sentryn with the active zone proteins SAD-1 and UNC-10 (RIM) in the DA9 motor neuron synaptic region. Sentryn-Emerald and SAD-1-tdTomato are coexpressed from the ceEx510 transgenic array. tdTomato-Sentryn and UNC-10 [627-974]-GFP are coexpressed from the ceEx503 transgenic array. (B) Representative images showing localization of Sentryn-GFP to the ventral apex of boutons (the location of the active zone) at DB7 motor neuron synapses. Note the highly focal localization of Sentryn-GFP to the ventral apex as indicated with arrows. Single bouton images (indicated) are from three different animals, and includes two of the boutons enlarged from the multi-bouton image. Dashed lines outline the approximate boundaries of the boutons as determined by the nonfocal Sentryn-GFP signal that fills the bouton at a lower concentration. Sentryn-GFP is expressed from the ceEx462 transgenic array, and is functional because it restores wild-type function to a strn-1 null mutant (Figure S4). (C) Representative images showing localization of Sentryn-GFP in the DB7 motor neuron synaptic region in the indicated mutant backgrounds. The single- and multi-bouton images were all acquired from different animals. Note that the highly focal localization of Sentryn-GFP is greatly disrupted in the syd-2 null mutant. The scaling of these images has been altered by ∼50% to brighten the images for viewing the nonfocal localization. Sentryn appears to fill boutons, but is no longer enriched at the ventral apex where the active zone is located. As shown in the single bouton enlargements, Sentryn sometimes shows a patchy diffuse localization in syd-2 mutant boutons. In sad-1 mutant boutons, Sentryn localization appears partially disrupted, sometimes localizing to the ventral apex and sometimes appearing unlocalized or localizing to bouton regions other than the ventral apex. In general, Sentryn localization is variable in mutants lacking sad-1. To illustrate the scope of the variability, a panel of images of Sentryn localization in the sad-1 null mutant is shown in Figure S6.