Cholera toxin and wheat germ agglutinin conjugates as neuroanatomical probes: Their uptake and clearance, transganglionic and retrograde transport and sensitivity

Abstract

Horseradish peroxidase (HRP) conjugates of 6 different lectins and cholera toxin (CTHRP) were quantitatively compared with respect to: (a) their behavior at the injection site and (b) their ability to label, by means of transganglionic and retrograde axonal transport, axon terminals and neurons in the medulla of the rat subsequent to injections of each probe into the anterior two thirds of the tongue. HRP conjugates of wheat germ agglutinin (WGHRP) and CTHRP were more sensitive than any of the other lectin-HRP conjugates. Both were far superior to free-HRP (FHRP) in demonstrating these projections and CTHRP was the most sensitive transganglionic and retrograde probe. Additional experiments demonstrated that this superiority was not an artifact of the volume of material injected into the tongue nor of the injection site area or survival time selection. These experiments demonstrated further that CTHRP and WGHRP remain at the injection site approximately twice as long as FHRP and that their removal from or degradation in retrogradely labeled neurons requires approximately twice as much time as that required for FHRP. These observations, together with earlier studies from this laboratory, suggest the following conclusions: (1) CTHRP and WGHRP are superior in sensitivity to FHRP for studies of neuronal connectivity; and (2) HRP conjugates of ligands such as CTHRP and WGHRP are internalized, transported and/or degraded by mammalian neurons in a manner which differs from that of FHRP, a macromolecule for which neuronal plasma membrane ‘receptors’ are lacking.