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Fig. 5. Apical polarity of spermatheca cells is affected by par-3(RNAi). (A-D)
Mislocalization of AJM-1::GFP in spermathecae of RNAi-treated worms. Worms
carrying AJM-1::GFP were collected at 39 hours after growth with (right) or
without (left) RNAi treatment and localization of AJM-1::GFP and anti-LET-413
was analyzed by confocal microscopy. The distal spermatheca is to the right in
all panels; the distal-most cell pairs are indicated by brackets in (C) and
(D). Panels (A) and (B) show projections of 36 and 26 optical sections through
the entire spermatheca, respectively. (C,D) Single optical sections selected
to show the middle focal plane through the distal spermatheca.
(C',D') Summary of our interpretation of the observed defects in
the distal spermatheca. In control worms, AJM-1::GFP is precisely localized to
the apical junctions of the distal spermathecal tube (arrowheads). However, in
par-3(RNAi) worms, AJM-1::GFP is widely dispersed in puncta that extend into
the lateral, and in some worms into the basal, domains. Gaps of GFP signals
(arrowheads) are often observed. (E,F) Mislocalization of microfilaments in
PAR-3-depleted spermatheca. Worms carrying LET-413::GFP were collected after
48 hours of growth with (right) or without (left) RNAi treatment and stained
with rhodamine-phalloidin (red in top panels) after dissection. Ovary (ov) and
the spermatheca (sp) are indicated. In control worms, microfilaments are
concentrated beneath the apical cell membrane in cells near the distal end of
the spermathecal tube (left column, arrow). However, in par-3(RNAi) worms,
microfilaments are lacking in this region (right column, arrow). Note that
although cell shape is abnormal, LET-413::GFP remains restricted to the
basolateral membrane domain (arrowheads). Scale bar: 8 µm (A-D); 5 µm
(E-F'').
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