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Fig. 6. Distribution of FER-1 and 1CB4 in spermatids and spermatozoa. (A,B) Ultrastructural localization of FER-1 with AZ10 antibodies in wild-type (A) spermatids and (B) spermatozoa. (C,D) Localization of 1CB4 staining in wild-type (C) spermatids and (D) spermatozoa. MO, membranous organelles; M, mitochondrion; LM, laminar membranes; P, pseudopod. Arrowhead indicates MO head; double arrowhead indicates MO body. Bar, 500 nm. (E-H) Immunogold label was quantitated over cellular compartments (E,F) and along membranes (G,H), for AZ10 (E,G) and 1CB4 (F,H). AZ10 labels MOs at the highest density (E), which decreases in the MO membrane and increases in the PM after fusion (G). 1CB4 labels MOs most abundantly (F), which increases two-fold in density along MO membranes and simultaneously increases slightly in the PM after fusion (H). Insets in E and F show average total labeling/cell for spermatids (tid) or spermatozoa (zoa) is not statistically different for either antibody. Insets in G and H show labeling over cell body (CB) or pseudopod (P) plasma membranes, which are not statistically different for either antibody. Graph indicates the average labeling over entire PM. Although nuclear localization was observed at a density similar to that of the MO by immunogold staining with AZ10, we observed no nuclear staining by immunofluorescence and presume this is background staining due to the charged nature of the nucleic acids. Error bars=s.e.m. ^*Different from unfused body density (P<0.0001); ^**different from cell body cytoplasm density (P<0.0001); + different from unfused body linear density (P<0.0001); ++ different from spermatid PM linear density (P<0.0001).

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