FHOD-1::GFP is localized in F-actin rich muscles, including BWMs, pharyngeal muscles, and vulval muscles. FHOD-1 is present diffusely in the elongated bands of F-actin rich embryonic myoblasts. FHOD-1 is not detectable in F-actin rich BWMs of L1 larvae but appears as puncta along muscle edges and as striations across the BWM contractile lattice in L2 and L3 larvae.
Immunoreactivity was concentrated in nervous system regions rich in synapses, including the nerve ring, the dorsal nerve cord and the ventral nerve cord.
The antibody specifically stained the synapse-rich nerve ring, ventral nerve cord and dorsal nerve cord in wild-type but not in unc-43(n1186) or unc-43(e755) animals.
Rim staining was restricted to discrete puncta in synapse-rich regions of the nervous system including the nerve ring, the ventral nerve cord and the dorsal nerve cord. Localized Rim protein was observed in all larval stages.
On whole-mount in situ preparations, the affinity-purified anti-RPM-1 antibodies detected weak expression of endogenous RPM-1 in the nerve ring, a synapse-rich region, in wild-type animals but not in rpm-1(ju23) and rpm-1(ju44) mutants.
EHS-1 was concentrated in synaptic-rich structures of the worm nervous system: the nerve ring and its associated ganglia; and the ventral and dorsal cord processes.The staining of the two cord processes appeared punctate and organized in an almost continuous line. Immunoreactive dots in the ventral and dorsal sublateral processes were smaller than those in the main cords. No detectable staining was observed in neuronal cell bodies, commissures or any other tissue.
CYK-1 is expressed in the excretory cell (EC) using translational green fluorescent protein (GFP) fusions expressed from fosmids (Sarov et al., 2012). CYK-1::GFP is found in the cytoplasm and in punctate structures in many tissues. In the EC it accumulates at the actin-rich leading edge and in the cytoplasm. Localization of a functional Venus::CYK-1 specifically expressed in the EC displays similar accumulation, with visible punctate structures that also accumulate LifeAct::TagRFP.
We observed FHOD-1::GFP in BWMs of animals from embryonic 1.75-fold stage through L1. FHOD-1 localized diffusely in younger stage embryonic BWM when sarcomeric components start to polarize and assemble into sarcomeres, and gradually appeared more punctuate at the edges of the F-actin-rich contractile lattices from later embryogenesis through early larval development, when sarcomeres mature. Similar patterns of fluorescence were visible in live embryos and larvae expressing FHOD-1::GFP. Thus, FHOD-1 is present through all stages of BWM development when sarcomere assembly is occurring.