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Fig. 3. Bam promotes differentiation of TkvAct ovarioles in part by downregulating
Dpp signaling prior to overt GSC differentiation. (A-C) Fluorescence (A) and
confocal (A inset, B,C) projections of ovarioles from females of genotype
P{hs-bam.O}/+;
P{UAS.p-TkvAct)/P{Gal4::VP16-nos.UTR}. Anti-Orb (red). (A) DAPI
staining (green). Ovariole of female subject to daily heat shock from day 10
of development. (Inset) Abnormal egg chamber from an ovariole of a similarly
treated female. (B,C) Yopro-1 staining (green) of (B) ovarioles and (C)
germarium of 10-day-old female subject to daily heat-shock from day 9-10 of
development until 7 days post-eclosion. Note complete rescue of the tumorous
phenotype (B), with differentiation of wild-type appearing egg chambers with
15 polyploid nurse cells and localization of Orb to the presumptive oocyte,
and wild-type appearance of the germarium (C). (D-K) Confocal projections
(D-G) or confocal sections (H-K) of germaria or ovarioles from females of
genotype P{hs-bam.O}/+; P{nos-myc.V}/+; P{UAS.p-TkvAct}
P{lacZ}DadP1883/P{GAL4::VP16-nos.UTR}. (D,E)
Anti-spectrin (red), anti-Myc (green). In a non-heat shocked control germarium
(D), all germ cells express Nos (circle, one example). In a germarium from a
2-day-old adult female heat shocked daily from day 9-10 of development (E),
all spectrosome-containing cells express Nos (circle, one example). Nos is
downregulated in all germline cysts that have between two and eight cells
(asterisks), but is re-expressed at high levels in 16 cell cysts (arrowhead).
(F,G) Anti-Myc (red), anti-lacZ (green). Ovarioles from 2-day-old
female without heat shock (F), or with daily heat shocks starting at day 10 of
development (G). In heat-shocked ovarioles, the level of Nos-Myc is unchanged
in germ cells, while the level of Dad-lacZ expression is
significantly reduced in germ cells, but not in follicle cells. (H)
Anti-lacZ (green). A confocal section from part of the projection in
G, showing lacZ expression is reduced but not eliminated in the
germline. (I-K) Germaria from control (J) and heat shocked (I,K) females
(anti-lacZ, green; anti-Hts, red). Compared with control females (J),
heat-shock causes reduction, but not elimination of lacZ expression
in spectrosome containing cells (K, asterisk), and complete elimination of
lacZ expression in fusome-containing cysts (I, asterisk). (L,M)
Confocal projections of ovarioles from females of genotype
P{UAS-lacZ.p}/P{hs-bam.O};
bamD86/P{GAL4::VP16-nos.UTR} bamD86
(anti-Hts, red; anti-lacZ, green). Germarium of non-heat shocked
control (L) and germarium of 2-day-old female heat shocked daily from day 9-10
of development (M), showing rescue of tumorous phenotype as evidenced by
differentiating egg chambers. Levels of lacZ are not reduced after
heat shock. (A-E,I-M) Arrow indicates terminal filament.
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