Figure 6. Immunocytochemical analysis of whole-mount worms using anti-Bpat-3cyto. Animals are oriented with the head to the left, dorsal up. (A)
bpat-3 staining in two quadrants of body wall muscles. Arrows indicate staining localized to the longitudinal cell boundaries. (B) Enlarged view of body wall muscle cells showing
bpat-3 staining localized to the dense bodies (arrows)and M lines (arrowheads). (C) Lateral view, showing
pat-3 staining of the ventral attachment sites (apposed to the anus) and the middle strut of the anal depressor muscle (ADM), the ventral attachment sites of the sphincter muscle (Sph), the basal surface of the intestinal muscles (IM), and the cell surface of the left postembryonically derived coelomocyte (CC).
bpat-3 staining in the dorsal and ventral body wall muscle quadrants is out of the plane of focus. (D) Lateral view, left side, showing
bpat-3 staining at the ventral and lateral attachment sites of the
vm1 and
vm2 vulval muscles (
vm1and
vm2), the anterior spermatheca (Sp), and the embryonically derived coelomocytes (CC). (E) Lateral view, right side, showing
bpat-3 staining in the vulval muscles as described in D, and staining at the lateral ridge attachment sites of the uml uterine muscles (uml), in the anterior spermatheca and in the posterior uterus (Ut) of a young adult hermaphrodite. Intestinal and body wall muscle staining are out of focus. (F) Nomarski image of the same animal as in E. (G) Competition of anti-
bpat-3cyto with 10 ug/ml of the cytoplasmic peptide. To ensure that the animals had been permeabilized, the monoclonal antibody MH27, which recognizes a desmosomal component at the apical surface of hypodermal, pharyngeal, and intestinal tract cells, was included in the sample. No
bpat-3 staining was observed. The pattern of MH27 staining is shown for comparison with H; the pharynx (ph) and the intestine (int) are indicated. (H) Staining of
pat-3(
rh54) embryos with anti
bpat-3cyto show that staining in the intestine (int) is not dependent on the expression of
bpat-3. Animals were colabeled with MH27 to provide an outline of the intestine; the pattern of MH 27 staining can be seen in G. Bars, 20 um.