A hypothesis for the superior sensitivity of wheat germ agglutinin as a neuroanatomical probe.
We have shown that a conjugate of wheat germ agglutinin (WGA), with horseradish peroxidase (HRP), is more sensitive than native HRP as a probe of neuroanatomic connections involving the retrograde transport of the lectin. It has also been shown in our laboratory that WGA-HRP remains at the site of injection twice as long as HRP. The purpose of the present morphometric study was to investigate the basis for the higher sensitivity of WGA-HRP over HRP as a retrogradely transported tracer molecule. To do this, we modified the experiment of Heuser and Reese which utilized the tracing of HRP in the frog neuromuscular junction (Heuser, J.E. and Reese, T.S., J. Cell Biol., 57 (1973) 315-344). Instead of using HRP alone, we examined, in double labeling experiments, fluid and adsorptive endocytosis with free HRP and WGA coupled to ferritin (WGA-ferritin) respectively. Immediately after nerve stimulation, both markers are taken up simultaneously into cisternae, and in tubular structures strikingly similar to the described compartment of uncoupling of receptor from ligand (CURL). Frequently, cisternae were connected with putative CURL. This early double labeling of cisternae and putative CURL was followed by the appearance of synaptic vesicles labeled with WGA-ferritin only (72-79%), HRP only (6-11%), and both labels (13-16%). In contrast to the labeling pattern of synaptic vesicles, the majority of cisternae and putative CURL had both labels throughout the duration of the experiments (77-80%). The results of this study indicate that most of WGA-ferritin and HRP are co-localized in cisternae and putative CURL, compartments involved in endocytosis and surface receptor recycling.(ABSTRACT TRUNCATED AT 250 WORDS)
Stieber
A
A
Division of Neuropathology, University of Pennsylvania, Philadelphia 19104.
Erulkar
S D
SD
Gonatas
N K
NK
D000818:Animals; D004705:Endocytosis; D006735:Horseradish Peroxidase; D009046:Motor Neurons / Q000166:cytology* / Q000502:physiology; D009434:Neural Pathways; D010544:Peroxidases; D011894:Rana pipiens; D014909:Wheat Germ Agglutinins / Q000493:pharmacokinetics
D014909:Wheat Germ Agglutinins; D006735:Horseradish Peroxidase; D010544:Peroxidases