Chapter 2 32 representation of peak fluorescence intensities in the retinal vascular bed of both genotypes. (d-i) Histopathological functional evaluation: (d) A representative proximal tubule of wt larva injected with the 70-kDa labelled dextran(bar=10 µm). (e) A representative proximal tubule of wt larva injected with the 4-kDa labelled dextran (bar=10 µm). (f) A representative proximal tubule of ctns‒/‒ larva injected with the 70-kDa labelled dextran(bar=10 µm).(g) A representative proximal tubule of ctns‒/‒ larva injected with the 4-kDa labelled dextran (bar=10 µm). (h) A higher magnification of the proximal tubules of both genotypes showing internalized 70-kDa dextran within cytosolic puncta that likely correspond to endocytic compartments (marked areas in panels d and f) (bars=5 µm). (i) Quantitation of the number of dextran puncta in both high and low molecular weight dextran injections in both genotypes (N=10 for each genotype and each condition). * P<0.05, *** P<0.001. Figure 7. Megalin expression in proximal tubular cells. (a) Transverse fluorescent image of the proximal pronephric region of wt 5 dpf larva labelled with anti-megalin antibody (bar=10 µm). (b) Higher magnification image of wt proximal tubule (square in panel a) showing mainly the diffuse distribution of megalin at the cellular brush border (bar=3 µm). (c) The proximal pronephric region of ctns‒/‒ 5 dpf larva labelled with anti-megalin antibody (bar=10 µm). (d) Higher magnification image of ctns‒/‒ proximal tubule (square in panel c) showing majority of megalin staining in sub-apical intracytoplasmic vacuoles (white arrows) (bar=3 µm). Outer boundaries of proximal tubules were delineated with green, lumen with red, and nuclear boundaries were delineated with white. (e) Quantitation of megalin protein abundance in proximal tubules of wt and ctns‒/‒larvae (N=5 for each genotype). (f) Quantitation of the megalin encoding lrp2a RNA expression in homogenized larvae of 6 dpf wt vs ctns‒/‒larvae (N= 5 individually separated RNA samples for each genotype). * P<0.05. Figure 6. Continued
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