We are characterizing the body wall muscle phenotype of
unc-78 mutants in C.elegans. Waterston, et al. (Dev. Bio.77:271,1980) described the phenotype of
unc-78 mutants as disruption of the myofilament lattice with accumulation of thin filaments at the periphery of body wall muscle cells. These animals are only slightly slower and uncoordinated when compared to N2. There are three known alleles of
unc-78: CB1217 (reference), CB1221, and ST43. Fluorescence microscopy images of phalloidin stained animals reveals large aggregations of actin filaments in addition to normal looking thin filaments. Immunofluorescent images of CB1217, using anti-Myosin A and anti-Myosin B, antibodies, reveals a discontinuous staining pattern for Myosin A with Myosin B staining appearing to be distorted in comparison to wild type. These results suggest that the disruption of the thin filament lattice also impacts the thick filaments. We have examined whole animal homogenates using SDS-PAGE and have analyzed those gels using quantitative densitometry. We determined protein accumulation profiles for the folloing: Histone H2A (internal standard), Tropomyosin, Actin, Paramyosin, and Myosin in both mixed population and L2/L3 synchronized populations. For two of the alleles: CB1217 and ST43, there are significantly increased actin levels in the body wall muscle. Tropomyosin and Paramyosin levels are not significantly impacted, while myosin levels are decreased, suggesting again that the disruption of the thin filament structure is also affecting the organization of myosin in the thick filaments. CB1221 is only mildly affected, with its protein profiles being very similar to N2. Analysis of
sup-13, the suppressor for
unc-78, reveals that the restoration of the wild type phenotype comes via significant elevation of myofilament proteins, most noticeably for myosin.