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Fig. 3. CEPsh glia support axon guidance in the nerve ring. (A-H) AWC
axonal defects of adult C. elegans lacking ventral left and right
CEPsh glia. Fluorescence images (A-D) and corresponding schematics (E-H) of
AWC neurons expressing odr-1::RFP. (A,E) Mock-ablated animal.
(B-D,F-H) AWC defects frequently observed in operated animals. In all panels,
the reporter is also expressed in AWB axons. (J-Q) AWC axon defects
observed in mls-2(ns156) adults. Fluorescence images (J-M)
and corresponding schematics (N-Q) of AWC neurons expressing
odr-1::RFP. (J,N) Wild-type AWC axon. (K-M,O-Q) AWC defects of
mls-2(ns156) adults. In J-M, reporter is also expressed in
AWB neurons. Similar defects are seen in vab-3(ns157)
mutants and in AFD neurons of mls-2(ns156) and
vab-3(ns157) animals. (I) Histogram depicting AWC,
ADF and AFD axonal defects in animals of indicated genotype or in which CEPsh
glia were ablated. VL/R+, expressing hlh-17::GFP in ventral left and
right CEPsh glia. Others, lacking hlh-17::GFP in one or both ventral
CEPsh glia. n, number of animals. Some mls-2(ns156)
mutants fail to express AWC reporters. We only scored animals in which robust
expression was evident. (R-U) DIC (R,T) and fluorescence (S,U) images
of wild-type L1 animal (R,S) and an L1 with ablated ventral left and right
CEPsh glia (T,U) expressing unc-119::GFP. The posterior pharyngeal
bulb is circled. Arrowhead, nerve ring. Note the abnormal positioning and
shape of the nerve ring in the ablated animal. (V) Mosaic studies of
mls-2. Axonal defect +, AFD axon defect; axonal defect -, no defect;
n, number of animals; transgene +, transgene present; transgene -,
transgene absent.
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-shaped lines, introns; arrow, position
of a previously predicted translation start site different from that described
here (see Materials and methods); TTTTCAG, trans-splicing site; the bHLH
domain is in blue. (F-N) Fluorescence (F,G,I,J,L,M) and merged
DIC/fluorescence (H,K,N) images of wild-type (F-H),
mls-2(ns156) (I-K) and vab-3(ns157) (L-N)
adults expressing hlh-17::GFP and ptr-10::myrRFP. White and
yellow arrowheads indicate dorsal and ventral CEPsh glia, respectively.
Asterisks indicate non-CEPsh glia. Scale bars: 5 µm.
)]. (E,H) Enlarged regions of D, (F,I) DIC and (G,J)
fluorescence images of a ventral right (E-G) and left (H-J) CEPsh glial cell
precursor (red arrowhead) in a 270-minute embryo expressing
mls-2::GFP. (K-N) Same as D-G but showing dorsal left and
right CEPsh glia precursors.
