NEUROFILAMENTS ARE PROMINENT IN BULLFROG OLFACTORY AXONS BUT ARE RARELY SEEN IN THOSE OF THE TIGER SALAMANDER, AMBYSTOMA-TIGRINUM

Bullfrog olfactory axons show variable numbers (0-29) of structurally typical neurofilaments (NFs) about 10 nm in diameter. In studies tracking these NFs through serial sections of axons in cross section, they were found to be discontinuous, with a calculated average length of about 118-mu-m. In contrast to olfactory axons in bullfrogs, those of the tiger salamander Ambystoma trigrinum rarely show NFs. To be certain that the absence of NFs is a specific characteristic of olfactory axons, pieces of salamander spinal cord, optic nerve, and sciatic nerve were examined and found to contain typical NFs. To minimize the possibility that NFs in salamander olfactory axons were degraded or poorly fixed during preparation for electron microscopy, samples were fixed by using a variety of fixative and buffer combinations. To exclude the possibility that proteases degraded NFs during processing, prior to fixation some pieces of olfactory nerve were incubated in physiological saline containing protease inhibitors. Regardless of the preparation method, NFs were generally not seen in salamander olfactory axons. Extracts of salamander olfactory nerve were subjected to SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblotting studies with monoclonal antibodies to the three NF subunit proteins. The immunoblots showed negligible or trace amounts of NF-L (light) and NF-H (heavy), while an NF-M (medium) protein having a molecular mass (Mr) of 160 kD was present in abundance. Extracts of salamander spinal cord, on the other hand, showed all three subunit proteins (with Mrs of 230, 160, and 77 kD). If one assumes that cells assemble structural elements to provide for a given function, the findings suggest that NFs in olfactory neurons of bullfrogs provide a function that may be missing in olfactory neurons of the salamander; the evidence also suggests that the absence of NFs in the salamander may be due to a deficiency in two of the three NF subunit proteins.