WormBase Tree Display for Picture: WBPicture0000009912

WBPicture0000009912 Description: Figure 2. GLD-3 Protein during C. elegans Development. (A) Two GLD-3 proteins. A Western blot of proteins prepared from adult animals that were wild-type (wt) hermaphrodites (lane 1), wild-type males (lane 2), glp-1 mutant hermaphrodites raised at restrictive temperature which lack a germline (no GL; lane 3), gld-3(q730) homozygous hermaphrodites (lane 4), and gld-3(q730); smg-1 homozygous hermaphrodites (lane 5), was probed with either of two GLD-3 antibodies: anti-GLD-3 (top blot), which detects both GLD-3L and GLD-3S, or anti-GLD-3L (middle blot). Actin was used as a loading control (lower blot).(B-M) Immunocytochemistry to determine location of GLD-3 protein. Samples shown were stained with anti-GLD-3 antibody. Similar results were obtained with anti-GLD-3L antibodies (data not shown). Epi-fluorescent images (B-H, L, and M); confocal images (I-K).(B and C) Wild-type L1. Gonadal primordium with the two germline precursors, Z2 and Z3, stained with anti-PGL-1 (B) and anti-GLD-3 (C). For both antibodies, antigen is cytoplasmic; the nucleus is seen as a dark area in the middle of each cell.(D-G) Extruded germlines of wild-type adult hermaphrodite (D and E) and male (F and G). DAPI (D and F); anti-GLD-3 (E and G). In both sexes, GLD-3 is predominantly cytoplasmic and present throughout the germline, except in secondary spermatocytes and mature sperm. Oocytes stain intensely with anti-GLD-3 (extent of oocytes marked by solid line; most proximal oocyte nucleus marked by small arrowhead). Primary spermatocytes also stain intensely (long arrowheads marked with 1); secondary spermatocytes (small arrow marked with 2) and mature sperm (extent marked by dotted line) have no detectable GLD-3. The level of GLD-3 is elevated, but not intense, in the distal germline. GLD-3 is more prominent in male pachytene than hermaphrodite pachytene germ cells, making it easier to visualize GLD-3 in granules surrounding the nuclear membrane (G). Insets in (D) and (E), 3 magnification showing GLD-3 in oocytes and primary spermatocytes. Insets in (F) and (G), 3 magnification showing early meiotic region from germlines of different males.(F) DAPI-stained nuclei.(G) Double stained with anti-PGL-1 (green) and anti-GLD-3 (red) antibodies. GLD-3 and PGL-1 overlap in most granules (yellow, arrow), but not all (red, closed arrowhead).(H-K) Wild-type 12-cell embryo.(L and M) Twelve-cell gld-3(q730) embryo from gld-3(q730) homozygous mother.(H) DAPI.(I and L) Anti-PGL-1.(J and M) Anti-GLD-3.(K) Merged image of (I) and (J) showing colocalization of GLD-3 with PGL-1 (yellow, arrows). In some granules, GLD-3 does not colocalize with PGL-1 (red, arrowhead). In gld-3(q730) mutant embryos that cleave, P granules are observed (L), but GLD-3 is absent (M).
  Name: F2.jpg
  Depict: Expr_pattern: Expr2269
    Anatomy: WBbt:0005784
    Cellular_component: GO:0005737
      GO:0043186
  Acknowledgment: Template: Reprinted from <Journal_URL>, <Article_URL>, Copyright <Publication_year>, with permission from <Publisher_URL>.
    Publication_year: 2002
    Article_URL: DOI id 10.1016/S1534-5807(02)00322-2
    Journal_URL: DevelopmentalCell
    Publisher_URL: Elsevier
  Reference: WBPaper00005566