Figure 6.

Zebrafish Embryos with gmppb Knockdown Have Morphological Defects, Damaged Muscle, and Hypoglycosylated α-DG at 48 hpf

(A) Bright-field microscopy of live embryos shows morphological defects of the gmppb-MO-injected embryos (injected with gmppb splice-blocking MO 3ng + p53 MO 6ng) as compared to uninjected wild-type embryos.

(B) Phalloidin staining of filamentous actin (red) and immunostaining with an antibody against β-DG (green).

(C) Live gmppb-MO-injected embryos injected with EBD (red) and imaged by confocal microscopy. Some fibers showed EBD infiltration, indicating damage to the sarcolemma (green arrow), and other fibers detached from the myosepta and retracted (yellow arrow) and thus left a space. The following abbreviation is used: DIC, differential interference contrast. The scale bar represents 25 μm.

(D) gmppb morphants have significantly more interfiber spaces than do wild-type uninjected embryos. The horizontal dotted line shows the median.

(E) An immunoblot shows a reduction in α-DG glycosylation. Percentage figures indicate the intensity of morphant bands relative to that of the wild-type and are adjusted for the γ-tubulin loading control. “gmppb MO” indicates embryos injected with gmppb MO 3ng + p53 MO 6ng, and “dag1 MO” indicates embryos injected with dag1 translation-blocking morpholino (5 ng).