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Analysis of glycolytic enzyme co-localization in Drosophila flight muscle

David T. Sullivan1,*, R. MacIntyre2, N. Fuda1, J. Fiori1, J. Barrilla1 and L. Ramizel1

1 Department of Biology, Syracuse University, Syracuse, NY 13224, USA
2 Department of Genetics and Molecular Biology, Cornell University, Ithaca, NY 14853, USA



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Fig. 1. Glycerol-3-phosphate dehydrogenase (GPDH) localization in antibody-stained myofibrils. Myofibrils were isolated from wild-type Drosophila muscles and reacted with anti-GPDH serum and then reacted with fluorescein isothiocyanate-labeled goat anti-rabbit serum and visualized as described in Materials and methods.

 


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Fig. 2. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and aldolase localization in antibody-stained myofibrils, as in Fig. 1.

 


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Fig. 3. Triose phosphate isomerase (TPI), phosphoglycerate kinase (PGK) and phosphoglycerol mutase (PGLYM) localization in antibody-stained myofibrils, as in Fig. 1.

 


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Fig. 4. A comparison between (A) phalloidin and (B) anti-glycerol-3-phosphate dehydrogenase (GPDH) doubly stained myofibrils. Note that the thin Z-discs and the broader M-lines in the phalloidin-stained myofibril coincide with fainter anti-GPDH stained and brighter stained corresponding bands, respectively.

 


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Fig. 5. Myofibrils from wild-type Drosophila muscles incubated for 5 min in contraction buffer and stained for glycerol-3-phosphate dehydrogenase (GPDH). Note the reduced staining at the M-lines as compared with Fig. 1.

 


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Fig. 6. Myofibrils from wild-type Drosophila muscles incubated in contraction buffer for 20 min and simultaneously stained using anti-glycerol-3-phosphate dehydrogenase (GPDH) and phalloidin. Note the correspondence of the brightly fluorescent GPDH signal with the very faint, narrow dark bands on the phalloidin-stained myofibril.

 


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Fig. 7. Glycerol-3-phosphate dehydrogenase (GPDH), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and aldolase in antibody-stained myofibrils prepared from GPDH null mutant flies. Visualized as in Fig. 1. The exposure has been extended so that an image of the weakly fluorescent myofibril would be evident.

 


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Fig. 8. Triose phosphate isomerase (TPI), phosphoglycerate kinase (PGK) and phosphoglycerol mutase (PGLYM) in antibody-stained myofibrils prepared from glycerol-3-phosphate dehydrogenase (GPDH) null mutant flies. Visualized as in Fig. 1. The exposure has been extended so that an image of weakly fluorescent myofibril would be evident.

 





© The Company of Biologists Ltd 2003