Ribosomes and polyribosomes are present in the squid giant axon: an immunocytochemical study.
Ribosomes and polyribosomes were detected by immuno-electron microscopy in the giant axon and small axons of the squid using a polyclonal antibody against rat brain ribosomes. The ribosomal fraction used as antigen was purified by ultracentrifugation on a sucrose density gradient and shown to contain ribosomal RNAs and native ribosomes. The polyclonal antibody raised in rabbits reacted with at least ten proteins on immunoblots of purified rat brain ribosomes as well as with a set of multiple ribosomal proteins prepared from the squid giant fiber lobe. Immunoreactions were performed on cryostat sections of the stellate nerve cut at a distance of more than 3 cm from the stellate ganglion, using pre-embedding techniques. Ribosomes and polyribosomes were identified within the giant axon and small axons using electron microscopic methods, following binding of peroxidase-conjugated anti-rabbit IgG secondary antibody. Polysomes were more frequently localized in peripheral axoplasm, including the cortical layer of the giant axon, and were generally associated with unidentified cytoskeletal filaments or with dense matrix material. The immunochemical demonstration of ribosomes and polyribosomes in the giant axon and small axons of the squid confirms similar observations in the squid and the goldfish obtained with the method of electron spectroscopic imaging, and strongly supports the view that a local system of protein synthesis is present in axons. The immunochemical method here described offers an alternative tool for the selective identification of ribosomes, and is likely to prove of value in the analyses of other axonal systems.
Sotelo
J R
JR
División Biofísica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.
Kun
A
A
Benech
J C
JC
Giuditta
A
A
Morillas
J
J
Benech
C R
CR
D000818:Animals; D000906:Antibodies; D001369:Axons / Q000648:ultrastructure*; D001921:Brain / Q000648:ultrastructure; D002458:Cell Fractionation; D049832:Decapodiformes; D017952:Ganglia, Invertebrate / Q000166:cytology / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D009412:Nerve Fibers / Q000648:ultrastructure; D009474:Neurons / Q000648:ultrastructure; D011132:Polyribosomes / Q000648:ultrastructure*; D012335:RNA, Ribosomal / Q000032:analysis; D011817:Rabbits; D051381:Rats; D012269:Ribosomal Proteins / Q000032:analysis; D012270:Ribosomes / Q000648:ultrastructure*
D000906:Antibodies; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins