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Fig. 3. Expression of NHE3, Na+/K+-ATPase and V-ATPase in the
gastric caeca of larval Aedes aegypti. AeNHE3 is highly expressed in
the distal segment of gastric caeca (A) predominantly in the basolateral
membrane (orange in B) of distal segment. Only faint labeling of NHE3 was
found in the proximal segment of caeca (C, arrow).
Na+/K+-ATPase is excluded from regions of higher NHE3
expression but is enriched in proximal segment (yellow in E) and is localized
to the basolateral membrane (yellow in F). V-ATPase localizes to the apical
plasma membrane of both proximal and distal parts of the gastric caeca (H,I).
Weaker V-ATPase labeling was found in the distal basal membrane (arrow in H).
Gastric caeca was labeled with anti-NHE3 antibody alone (A-C) (red in A,
orange in B,C) or together (red in E-G) with
anti-Na+/K+-ATPase monoclonal antibody (yellow in E-G).
To visualize structures, actin was labeled with Alexa Fluor 488-phalloidin in
B-D (green). Expression of the V-ATPase was detected with polyclonal
antibodies to the B-subunit (H,I). Preimmune serum was used as negative
control (D) with phalloidin (green). Immunohistochemistry was performed on
whole mounts (A,E) or tissue sections as described in Materials and methods.
E1 (red for NHE3) and E2 (yellow for Na+/K+-ATPase) show
the separate channels that are merged in E. F and G are similarly merged
images of dual labeling with NHE 3 (red) and
Na+/K+-ATPase (yellow) antibodies. Scale bars, 100 µm
(E,E1,E2); 50 µm (A-D,F-I).
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