Representative confocal z-projection images of amphid (head) and phasmid (tail) neurons from hermaphrodites of the indicated genotype expressing GFP::RAB-28Q95L. Anterior is to the left; all images …
(A) Representative confocal z-projection images of amphid and phasmid neurons from worms of the indicated genotypes expressing GFP::RAB-28Q95L. Images are the same as those in Figure 1 except image …
Data for Figure 1 and Figure 1—figure supplement 1B.
(A) Epifluorescence z-projections of the heads and tails of C. elegans males expressing rab-28p::sfGFP and klp-6p::tdTomato (EVN cilia-specific reporter). Arrows denote the CEM neurons. Brightfield …
Data for Figure 2C.
(A) Transmission electron microscopy (TEM) images showing the amphid channels of the indicated genotype in cross section, at the positions of ciliary distal segments (DS) and middle segments …
(A) Left- schematic showing the location of the deleted regions in the gk1040 and tm2636 mutant alleles of rab-28. Right- cartoon modified from Bae et al. (2006) depicting the morphology of the CEM …
Data for 4C-E, 4G-I, 4 K-N.
Top schematic shows the location of the deleted region in the tm2636 mutant allele and the location of the primer pairs used to amplify full length cDNA sequence that was sequenced using the …
(A) Fluorescence micrographs of control and rab-28(gk1040) adult males expressing CIL-7::GFP. Close-ups of the male heads are shown on the left. Dotted white boxes mark one cilium. White arrowheads …
Data for Figure 4—figure supplement 2B,C.
(A) Cartoon of the ultrastructure of the cephalic sensory organ reproduced from Wang et al. (2014a). EVs (magenta spheres) are ‘shed’ from the PCMC/ciliary base into the lumen and ‘released’ into …
(A, E) Cartoon of the ultrastructure of the cephalic sensory organ reproduced from Wang et al. (2014a). Color scheme is same as that in Figure 5. The distal and proximal cephalic lumens (A) and PCMC …
Data for Figure 5—figure supplement 1B,C,F.
(A) TEM cross sections of control, rab-28(tm2636), and bbs-8 amphid channel cilia at the middle segment (top) and at the transition zone (bottom). Yellow arrowheads in rab-28 indicate the occasional …
Left- cartoon representation of the amphid sensory organ with the locations of the different ciliary segments marked. Right-Transmission electron micrographs of chemically frozen control (N2) adult …
Color scheme is the same as in Figure 5; magenta spheres represent shed EVs accumulating within sensory organs and EVs released into the environment, brown spheres represent secreted molecules …
BBS-8:: GFP levels in rab-28(tm2636) are slightly reduced in ray neurons but, remain unaltered in amphid and phasmid cilia. Scale bar is 1μm for amphid and phasmid panels and 5μm for ray neuron cilia.
Anterior is to the left. Movies are played at 5 frames per second. Scale bars; 2 μm. PCMC: periciliary membrane compartment.
A RAB-28-positive IFT particle can be seen in the bottom CEM and RnB cilia. Higher frequency IFT movement of RAB-28 is evident from the amphid cilia. PCMCs are marked by asterisks. Movies are played …
Models depict the CEM cilium (gold), and EVs (magenta spheres). Dotted white line in movie shows position of the TZ. rab-28 mutants ectopically accumulate excess EVs in distal regions of the …
Amphid sensory organ TEM phenotypes | |||||
---|---|---|---|---|---|
Mutant | Sex | Sensory compartment size | Matrix filled vesicles in sheath | EVs in pore | Reference |
rab-28(tm2636) | Male | Not enlarged | Yes | Few vesicles/odd shaped particles observed surrounding MS | This work |
rab-28(gk1040) | Hermaphrodite | Not enlarged | No | ND | Jensen et al., 2016 |
RAB-28Q95L | Hermaphrodite | Enlarged | No | ND | Jensen et al., 2016 |
RAB-28T49N | Hermaphrodite | Not enlarged | Yes | ND | Jensen et al., 2016 |
bbs-8 | Male and hermaphrodite | Enlarged | Yes | Lots of EVs (in male) | This work |
arl-6 | Hermaphrodite | Not enlarged | Yes | ND | This work |
Male cephalic sensory organ TEM phenotypes | |||||
Mutant | Sensory organ size | EVs in pore | Reference | ||
rab-28(tm2636) | Enlarged | Yes, especially at the distal regions | This work | ||
rab-28(gk1040) | ND | ND | N/A | ||
RAB-28Q95L | ND | ND | N/A | ||
RAB-28T49N | ND | ND | N/A | ||
bbs-8 | Enlarged | Yes, surrounding all segments | This work | ||
arl-6 | ND | ND | N/A | ||
Other phenotypes | |||||
Phenotype | rab-28(tm2636) | rab-28(gk1040) | arl-6 | bbs-8 | |
PKD-2 localization in CEMs | Mislocalized | Normal (Jensen et al., 2016) | ND | ND | |
CIL-7 localization in CEMs | Mislocalized | Mislocalized | Mislocalized | Mislocalized | |
Environmental release of EVs from heads | Normal | Normal | Normal | Normal |
Key resources table.
Primers used in this study.
Means and SEM for all figures.