Expr15724

The pattern of DEB-1 localization correlated with the known pattern, i.e. targeting DEB-1 within the muscle dense bodies and muscle-like fibers in the somatic gonad cells, as reported by Ono et al. Additionally, we observed a reproducible signal within the gonadal distal part, namely in the cytoplasm and the nuclear area of meiotic cells. More interestingly, we identified strong and reproducible labeling in meiotic cells using a polyclonal antibody against DEB-1/b*f. After a detailed investigation of DEB-1/b*f isoform localization in the cytosol, we also investigated its nuclear localization. To the best of author ́s knowledge, this is the first observation of its kind. We used structured illumination microscopy (SIM; Nikon) with lateral resolution around 100 nm and identified DEB-1 inside the germ cell nucleus, partially colocalizing with DNA of the transition zone as well as pachytene nuclei. Transmission electron microscopy (TEM) analysis of DEB-1/b*f localization in germ-line nuclei confirmed localization to the early prophase I - in both leptotene/zygotene and pachytene stages. During leptotene/zygotene, DEB-1/b*f is localized in clusters in close vicinity of the inner nuclear membrane. Moreover, in the mid-pachytene regions of wild-type worms, DEB-1 localizes to the vicinity of the axial elements comprising the SC.

Expr15723

The pattern of DEB-1 localization correlated with the known pattern, i.e. targeting DEB-1 within the muscle dense bodies and muscle-like fibers in the somatic gonad cells, as reported by Ono et al. Additionally, we observed a reproducible signal within the gonadal distal part, namely in the cytoplasm and the nuclear area of meiotic cells. More interestingly, we identified strong and reproducible labeling in meiotic cells using a polyclonal antibody against DEB-1/b*f. After a detailed investigation of DEB-1/b*f isoform localization in the cytosol, we also investigated its nuclear localization. To the best of author ́s knowledge, this is the first observation of its kind. We used structured illumination microscopy (SIM; Nikon) with lateral resolution around 100 nm and identified DEB-1 inside the germ cell nucleus, partially colocalizing with DNA of the transition zone as well as pachytene nuclei. Transmission electron microscopy (TEM) analysis of DEB-1/b*f localization in germ-line nuclei confirmed localization to the early prophase I - in both leptotene/zygotene and pachytene stages. During leptotene/zygotene, DEB-1/b*f is localized in clusters in close vicinity of the inner nuclear membrane. Moreover, in the mid-pachytene regions of wild-type worms, DEB-1 localizes to the vicinity of the axial elements comprising the SC.