WormBase Tree Display for Expr_pattern: Expr2286

Expr2286 Expression_of: Gene: WBGene00006436
    Reflects_endogenous_expression_of: WBGene00006436
    Sequence: Y38B5A
  Expression_data: Life_stage: WBls:0000024
      WBls:0000038
      WBls:0000027
      WBls:0000035
      WBls:0000041
      WBls:0000019
      WBls:0000020
      WBls:0000021
    Anatomy_term: WBbt:0003675 Certain
    GO_term: GO:0031674
  Subcellular_localization: As visualized with confocal immunofluorescence microscopy, EU102 stains the center of I-bands, between adjacent dense bodies. This was determined in three ways. When co-stained with antibodies to alpha-actinin, a major component of nematode dense bodies, EU102 staining appears to lie between, and does not overlap, the dense bodies. When co-stained with antibodies to actin, which gives a ladder-like pattern, EU102 appears to lie within the "rungs of the ladder". Finally, co-staining with antibodies to tropomyosin, that has been shown to localize to the outer portions of the I-bands in a linear pattern, showed that EU102 labels between the rows of tropomyosin staining.
    EU134 localizes to the I-band, but with a broader distribution than that obtained with EU102 antibodies. EU134 stains an area between dense bodies (overlapping with EU102 staining), as well as an area deeper into the I-band.
  Type: Antibody: In order to verify the localization obtained with the EU102 antibodies, the localization was determined using antibodies generated to another region of the Ce titin sequence. Antibodies (EU134) was raised to residues 10,24210,578, a region containing the two C-terminal Ig and last FnIII domains of the 1.2 MDa isoform. Except for the C-terminal-most 27 residues that are unique to the 1.2 MDa protein, this region is also part of the middle of the 2.2 MDa protein. To allow co-staining with the previous antibodies (EU102), the new immunogen was injected into a different animal species, the guinea pig.
      Rabbit polyclonal antibodies (EU102) to a 352-residue region (42154567) that contains Ig domains 18 and 19 and 150 amino acid residues beyond, and is just N-terminal to the PEVT/K region. On the basis of the Ce titin gene structure, it would be expected that EU102 would react with the 2.2 MDa and 1.2 MDa isoforms.
  Pattern: Indirect immunofluorescence with EU102, demonstrates expression of the two largest isoforms in all muscles except for the pharynx. The pattern of expression and localization of Ce titin was examined in embryonic and larval muscles. During the 1.5 and 2-fold stages no specific staining was observed. Then, first appearing during the 2.5-fold stage, Ce titin appears in puncta which seem to follow the myofibril tracts. This punctate staining persists in the 3-fold stage, but then staining all but disappears in L1 larvae. However, during the L2 or L3 stages, and continuing during the remainder of development, Ce titin staining is localized in the same dashed pattern as seen in adult muscle.
  Picture: WBPicture0000010194
    WBPicture0000010195
    WBPicture0000010196
    WBPicture0000010197
  Remark: Because the N-terminal 96 amino acid residues fused to GFP localizes to dense bodies, and because a major component of dense bodies is a-actinin, the localization of EU102 epitopes was determined in an alpha-actinin null mutant background. These animals, with the genotype atn-1(ok84), have normal motility and nearly normal muscle structure by polarized light microscopy. By EM, however, they have extra accumulations of actin near the ends of the muscle cells and, significantly, the dense bodies are shorter and more broadly based than normal. Staining of atn-1(ok84) muscle with EU102 is in a much broader pattern than wild-type and there is also loss of the "dashed" appearance. It is possible that in an alpha-actinin null background, the largest Ce titin isoforms are not properly anchored to the dense bodies. Sequence: Y38B5A.
  Reference: WBPaper00005546
  Antibody_info: WBAntibody00000570
    WBAntibody00000571