FIGURE

Fig. 2

ID
ZDB-FIG-091030-1
Publication
Sohn et al., 2009 - A role for nephrin, a renal protein, in vertebrate skeletal muscle cell fusion
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Fig. 2

Nephrin morpholino experiments in zebrafish reveal smaller muscles. (A) Light micrograph of variable phenotypes noted with nephrin morpholino in zebrafish embryos at 4 days post fertilization (dpf). Many of the embryos appear curved and shorter (red arrows) compared with control (black arrow). (B) Light micrograph of wild-type (top) mismatched (middle) and nephrin morpholino embryos (MO1) at 4 dpf. (C) RT-PCR analysis of mismatched and nephrin morpholino-injected embryos confirms the expected splicing defect leading to an in-frame deletion of the transmembrane domain, rendering a protein no longer able to anchor at the membrane; this is manifest by a decrease in RT-PCR product size (from 398 bp to 272 bp). Lane 1, molecular weight markers; lane 2, uninjected wild-type embryo; lane 3, 1.25 ng nephrin morpholino MO1; lane 4, 1.25 ng mismatched morpholino; lane 5, 2.5 ng nephrin morpholino MO1; lane 6, 2.5 ng mismatched morpholino. (D-I) Morphometric analyses performed on whole-mount embryos stained with anti β-dystroglycan antibody confirm that the myosepta to myosepta distance of embryos injected with nephrin morpholino MO1 (F, I) are smaller than those injected with mismatched morpholino (E, H) or wild-type uninjected embryo (D, G). (J) Bar graph of myosepta length measurements from wild-type, mismatched MO1 and nephrin morpholino MO1 injected embryos. Asterisk denotes a statistically significant P value by t test (P d 0.00000004). Individual measurements are listed in Table S1. Whole-mount control (K) and nephrin morpholino (L) embryos stained with anti-laminin (red) and anti-myosin heavy chain antibody (green); nuclei are stained in blue with DAPI. Nephrin morpholinos displayed shorter myosepta and presence of clustered nuclei (white arrows). (M-O) Whole-mount 1 dpf embryos are stained with anti-β-catenin antibody (red), and nuclei are stained in blue with DAPI. (M) Wild-type control; (N) control mismatched morpholino (MISMO1); (O) nephrin morpholino (MO1). Open arrows in (M) and (N) point to nuclei that appear regularly aligned, whereas arrows in (O) point to clustered nuclei.

Expression Data
Genes:
Antibodies:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Day 4

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Day 4

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Proc. Natl. Acad. Sci. USA