Functional Characterization of a Novel Class of Morantel-Sensitive Acetylcholine Receptors in Nematodes
Fig 5
Immunolocalization of Hco-ACR-26 and Hco-ACR-27 in H. contortus.
H. contortus L2 larval stage and adult males were fixed and incubated with affinity-purified antibodies raised against Hco-ACR-26 and Hco-ACR-27 subunits. Alexa 594- and Alexa 488- labeled secondary antibodies were used to determine the localization of Hco-ACR-26 (red) and Hco-ACR-27 (green) respectively. (A and B) Transmitted light and corresponding fluorescent apotome imaging performed on H. contortus L2 stage. Nematode’s nuclei were stained in blue using DAPI. Panel A shows a staining for Hco-ACR-26 and Hco-ACR-27 in striated cells of the nematode. Panel B shows that in a deeper focal section, Hco-ACR-26 and Hco-ACR-27 were also found to be expressed in another tissue potentially corresponding to the nerve ring (white arrows). (C, D and E) Confocal microscopy performed on L2 stage (C and E) and cross section of adult males (D). Body wall muscle cells were stained using primary antibodies raised against the myosin protein and further revealed with secondary Alexa 350-labeled antibodies (blue). (E) Negative control performed with anti-Hco-ACR-26 and anti-Hco-ACR-27 pre-adsorbed with their respective peptide antigen. For A, B, C and E anterior region is to the right. The scale bars correspond to 20 μm.