Fig. 5. Protein localization of ryanodine receptor, IP3 receptor and
thapsigargin-sensitive Ca2+-ATPase in Drosophila
melanogaster adults. Cryostat sections were processed as mentioned in
Materials and Methods, and then incubated with TX-R-BODIPY-ryanodine (1
µM), FL-Heparin (2 µM) and FL-BODIPY-thapsigargin (5 µM). Panel A
shows a sagital section of an adult male fly incubated with
TX-R-BODIPY-ryanodine. Staining is seen in many tissues derived from all three
germinal layers. Panel B is the same section as in A, but showing the signal
elicited with FL-Heparin staining; profuse labeling is also seen, as in A. In
both panels, intense labeling is seen in muscle (marked fm and lm in A) and in
the intestine (marked i in A). Adult male flies are approximately 2-3 mm long.
Panel C shows two images from the same section at 100x magnification.
The image on the left shows labeling of TX-R-BODIPY-ryanodine (red) and the
image on the right shows FL-BODIPY-thapsigargin (green) staining of a
horizontal section of the retina and optic lobe. Intense colocalization of
staining is seen in the neuropils of the optic lobe [lamina (l), medulla (m),
lobula and lobula plate (lp)], with lesser staining in the photoreceptor cells
and optic lobe neuronal cell bodies. As shown here, colocalization of
TX-R-BODIPY-ryanodine and FL-BODIPY-thapsigargin was also coincidental in all
adult tissues examined. Panel D shows a 1000x magnification of indirect
flight muscles stained with TX-R-BODIPY-ryanodine in longitudinal section,
where the striated pattern of labeling is evident, and panel D' shows a
cross-section of leg muscles also stained with TX-R-BODIPY-ryanodine. In both
cases, staining is cytoplasmic. Panel E shows a 1000x magnification of
intestinal cells of an adult marked with TX-R-BODIPY-ryanodine in their
cytoplasm. These sections are representative of 20 independent experiments.
Abbreviations: a, antenna; b, brain; fm, indirect flight muscles; g, gonad; i,
intestine; l, lamina; lm, leg muscle; lp, lobula and lobula plate (in the
section in C the lobula plate is directly underneath the lobula); m, medulla;
p, proboscis; r, retina and photoreceptor cells.