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First published online August 23, 2004
Journal of Experimental Biology 207, 3299-3305 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.01156
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A conserved domain of alkaline phosphatase expression in the Malpighian tubules of dipteran insects

Pablo Cabrero*, Valerie P. Pollock*, Shireen A. Davies and Julian A. T. Dow{dagger}

Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 6NU, UK



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Fig. 1. Comparison of cell numbers in the five Malpighian tubules of mosquitoes. Total, principal and stellate cell counts, and cell count in alkaline phosphatase domain, for the longest (1) to the shortest (5) tubule in each insect. Data are means ± S.E.M. (N=10) for Ae. aegypti (left) and An. stephensi (right).

 


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Fig. 2. Alkaline phosphatase domain in an intact adult D. melanogaster tubule pair: (A) control tubules subjected to alkaline phosphatase staining protocol without chromogenic substrate; the junction with the ureter is shown with an arrow; (B) DAPI-stained; (C) as in B but subjected to alkaline phosphatase staining protocol in the presence of NBT/BCIP chromogenic substrate; the junction with the ureter is shown with an arrow; (D) higher magnification to show predominantly apical localisation of staining. Scale bars, 100 µm for A-C, 10 µm for D.

 


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Fig. 3. Intact Ae. aegypti tubules. (A) Control tubules subjected to alkaline phosphatase staining protocol without chromogenic substrate; the junction with the gut is shown with an arrow; (B) as in A but subjected to alkaline phosphatase staining protocol in the presence of NBT/BCIP chromogenic substrate; the junction with the gut is shown with an arrow; (C) DAPI-stained; (D) as in C but also stained for alkaline phosphatase. Scale bars, 100 µm.

 


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Fig. 4. Intact An. stephensi tubules. (A) Control tubules subjected to alkaline phosphatase staining protocol without chromogenic substrate; the junction with the gut is shown with an arrow; (B) as in A but subjected to alkaline phosphatase staining protocol in the presence of NBT/BCIP chromogenic substrate; the junction with the gut is shown with an arrow; (C) DAPI-stained; (D) as in C but also stained for alkaline phosphatase; (E) high-magnification view of lightly stained lower tubule, revealing predominantly apical staining. Scale bars, 100 µm for A-D; 10 µm for E.

 


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Fig. 5. Intact G. morsitans tubules. (A) Control tubules subjected to alkaline phosphatase staining protocol without chromogenic substrate; the junction with the ureter is shown with an arrow; (B) as in A but subjected to alkaline phosphatase staining protocol in the presence of NBT/BCIP chromogenic substrate; the junction with the ureter is shown with an arrow; (C) high-power magnification of B; (D) as in C but different tubule preparation showing lower tubule/ureter boundaries and normally close association of tubules at the lower segment (arrows); (E) DAPI-stained; (F) as in E but also stained for alkaline phosphatase. Lower tubule segments remain closely intertwined in tubules shown in E and F. Scale bars, 100 µm.

 


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Fig. 6. Single intact S. gregaria tubule. (A) Control tubules subjected to alkaline phosphatase protocol without chromogenic substrate; (B) as in A but with NBT/BCIP chromogenic substrate; (C) DAPI-stained; (D) high-magnification image of lower tubule, without chromogenic substrate; (E) high-magnification image of B. Scale bars, 100 µm.

 


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Fig. 7. Taxonomy to genus of the insects used in this study. Taxonomy is that of NCBI (http://www.ncbi.nlm.nih.gov/Taxonomy/).

 





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