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Fig. 5. Eph is expressed on axons of the developing and adult MB. (A)
Diagram of MB development, showing the left hemisphere. Broken line indicates
the midline. MB neuroblasts are shown in yellow, the axonal projections of
neurons in black, 
'/ß' neurons in green and
/ß neurons in red (see text for details). (B-P) Eph
expression, as assayed by Ephrin-Fc binding, during MB development. A
high-power view of the left hemisphere of a late embryonic/early 1st instar
larval brain (B-D), stained with anti-Fas2 (B) and Ephrin-Fc binding (C). Eph
is present throughout the neuropil at this stage, including the MB
neurons and their bifurcated axons (D, dorsal lobe; M, medial lobe; Ped,
penduncle). (E-G) A 3rd instar larval MB from an individual carrying the
neuron driver 201Y-Gal4 driving the expression of
UAS-mCD8:GFP stained for GFP (E) and Ephrin-Fc binding (F). Eph is
present at low levels throughout the neurons and their bifurcated
axons. (H-J) An early pupal MB from an individual carrying the MB-specific
driver OK107-Gal4 driving the expression of UAS-mCD8:GFP
stained with anti-GFP (H) and Ephrin-Fc binding (I). Eph is expressed
throughout the MB, including '/ß' neurons (arrows).
(K-M) Late pupal stage MB stained for Fas2 (K) and Ephrin-Fc binding (L). Eph
expression is restricted to primarily /ß lobes, here identified by
double labeling with Fas2, with only low level Ephrin-Fc staining present
within the terminal region of the lobe (arrow in I). (N-P) An adult MB
double stained for Fas2 (N) and Ephrin-Fc (O). Eph expression is maintained
within the /ß lobes, with the highest levels of staining
restricted to the terminal regions of the lobe (arrow in O). Fas2
expression at this stage also labels the fan-shaped body (arrowhead in N)
lying directly beneath the medial lobes. (Q,R) A pupal MB from
an individual carrying the OK107-Gal4 driver plus
UAS-mCD8:GFP and UAS-Ephrin:myc stained for GFP (green) and
myc (red). Ephrin-myc is concentrated within MB cell bodies (arrows) and
absent from axons within the peduncle (Ped, arrowhead in N). (S) One
hemisphere of a late 3rd instar larval brain double labeled to visualize
Ephrin mRNA expression (blue) by in situ hybridization and anti-GFP
(brown) to detect mCD8:GFP driven by the MB-specific OK107-Gal4
driver. Ephrin RNA expression is widespread within the brain at this
time, but is detected within the MB neuron cell bodies, two clusters of which
are in the focal plane (arrows). Co-labeled MB cell bodies project axons
within the peduncle (arrowhead indicates the middle of peduncle), here labeled
by mCD8:GFP. Genotypes: (E-G) 201Y-Gal4/+; UASmCD8:GFP/+;
(H-J,S) OK107-Gal4/+;UAS-mCD8:GFP/+; (Q,R) OK107-Gal4/+;
UASmCD8:GFP/UAS-Ephrin:myc.
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