Specific N-glycans regulate an extracellular adhesion complex during dendrite patterning (2022)
Figure 1

Figure 1 AMAN-2/Golgi alpha-mannosidase II is required for PVD dendrite patterning.

Fluorescent images (left panels) and tracings (right panels) of PVD neurons of the indicated genotypes. PVD is visualized by the wdIs52 transgene. Primary (1º), secondary (2º), tertiary (3º), and quaternary (4º) dendrites are indicated, and the cell body is marked with an asterisk. Anterior is to the left and dorsal is up in all panels. Scale bars represent 20μm.

Schematic of the Menorin complex, including DMA-1/LRR-TM, SAX-7/L1CAM, MNR-1/Menorin, and LECT-2/Chondromodulin II, as well as the negative regulator KPC-1/Furin.

Genomic environs of aman-2. An N-terminal transmembrane domain (TMD) is indicated, encoding a type II transmembrane protein. All alpha-mannosidase II proteins contain a glycosyl hydrolase 38 domain. Four nonsense alleles and one deletion allele (tm1078) of aman-2 are denoted.

Quantification of the number of quaternary branches (indicated in schematic) in different aman-2 alleles and in wild type control animals. All loss of function alleles of aman-2 display a significant decrease in PVD quaternary branch number. Data are represented as mean ± SEM. Statistical significance was calculated using the Kruskal-Wallis test and is indicated (****p ≤ 0.0001). n = 15 for all genotypes and are biological replicates.

Quantification of the number of full tertiaries (marked by secondary and tertiary branches as shown in schematic) in the genotypes indicated. Data are represented as mean ± SEM. Statistical significance was calculated using the Kruskal-Wallis test and is indicated (****p ≤ 0.0001; ns = not significant). n = 15 for all genotypes and are biological replicates.

Fluorescent images and tracings of PVD in mnr-1(dz213) hypomorphic animals alone and in combination with an aman-2(gk248486) null mutant. The dz213 introduces a L135F missense mutation in the DUF2181 domain of MNR-1/Menorin. PVD is visualized by the dzIs53 transgene. The cell body is marked with an asterisk. Scale bars represent 20μm.

Quantification of the number of full tertiaries in genotypes indicated and traced in (F). The loss of aman-2 severely enhances the mnr-1(dz213) phenotype. Control indicates wild type control animals as shown in (A). Data are represented as mean ± SEM. Statistical significance was calculated using the Mann-Whitney test and is indicated (****p ≤ 0.0001; ns= not significant). n >23 for all genotypes and are biological replicates.

Fluorescent images and tracings of PVD in kpc-1(dz254) hypomorph animals alone and in combination with an aman-2(gk248486) null mutant. PVD is visualized by the dzIs53 transgene. White arrows indicate self-avoidance defects in tertiary branches. Red arrows show gaps between tertiary branches (no self-avoidance defects). The cell body is marked with an asterisk. Scale bars represent 20μm.

Quantification of the percent of self-avoidance defects in genotypes indicated and traced in (G). Control indicates wild type control animals as shown in (A). The loss of aman-2 suppresses defects in the kpc-1(dz254) phenotype. Data are represented as mean. Statistical significance was calculated using the Z-test and is indicated (****p ≤ 0.0001). n >15 for all genotypes and are biological replicates.