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.

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