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Fig. 5. Ectopic PKD-2 expression (Punc-119::PKD-2::GFP) reveals
cell-type specific factors required for PKD-2 localization.
(A,B) A wild-type hermaphrodite expressing
Punc-119::PKD-2::GFP. (A) PKD-2::GFP localizes to neuronal cell
bodies in the head, but GFP is excluded from ciliary endings. Small arrowheads
indicate the anatomical location of OLQ (outer labial quadrant) cilia in the
left panels. PKD-2::GFP is absent from cilia in the head sensillae of the
adult hermaphrodite. (B) In the hermaphrodite tail, PKD-2::GFP is retained in
cell bodies of many neurons including the ciliated phasmid sensory neurons.
Phasmid cell bodies were identified by their location (arrows); small
arrowhead indicates approximate anatomical location of phasmid cilia.
PKD-2::GFP is absent from cilia in the phasmids of the adult hermaphrodite.
(C,D) A wild-type male expressing PKD-2::GFP in every neuron.
(C) In the head, PKD-2::GFP is localized to the ciliary endings of CEMs (large
arrowheads in the left panels). (D) In the male tail, PKD-2::GFP is detected
in ciliated endings (large arrowhead, R3B cilium) of all B-type ray neurons,
except R6B (asterisk), which does not normally express the polycystins.
(E,F)Punc-119::PKD-2::GFP expression in the
unc-101(m1) mutant. Arrows indicate PKD-2::GFP dendritic expression
in polycystin-expressing neurons. (E) In the head, PKD-2::GFP is distributed
throughout male-specific CEM neurons. In all other neurons, PKD-2::GFP remains
in cell bodies. (F) In the tail, PKD-2::GFP is uniformly distributed only in
all B-type ray neurons (arrow) except R6B (asterisks). unc-101 males
are Mab (male abnormal) and often fail to develop all ray process. (G)
In the lov-1 mutant, PKD-2::GFP aggregates in CEM cell bodies
(arrows) but not in other neurons, and PKD-2::GFP is detectable in CEM cilia
(large arrowheads), which are identified by location and cell body morphology.
The inset shows one CEM cell body with PKD-2::GFP aggregates (compare with the
Fig. 3I inset). A subset of
z-series confocal sections is projected to generate the inset.
(H) PKD-2::GFP in lov-1 RnB ray neurons forms aggregates in
cell bodies (arrow), and the level of ciliary staining is reduced. Large
arrowhead points at R3B cilia with reduced PKD-2 expression. Scale bars: 10
µm. The anterior outlines of the worm are drawn by hand according to the
transmission images. (I) The percentage of ciliary localization of
Punc-119::PKD-2::GFP in non-native and native cells. The reason that
a small fraction (3-7%) of native cells do not localize PKD-2::GFP in native
cilia may be due to mosaicism of the extrachromosomal array. When PKD-2::GFP
is detected in R6B cilia, PKD-2::GFP staining is reduced and mislocalized to
the distal dendrite. In the hermaphrodite, there is no detectable ciliary
PKD-2::GFP expression in the head. Specifically, OLQ cilia were scored because
of their distinct shape and location. One-hundred and eight pairs of RnBs were
scored. For hermaphrodites (OLQs and phasmids), a total of 112 animals were
scored. There are four OLQ and phasmid neurons in each animal. WT, wild
type
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