Selective inhibition of neuronal protein synthesis by retrogradely transported ricin.
The ability of the lectin, ricinus communis agglutinin I (ricin120), to undergo retrograde axonal transport and cause degeneration of neuronal cell bodies has been frequently exploited to establish the origin of peripheral axons. Since this cytotoxic action of ricin results from its inactivation of ribosomes, the retrogradely transported lectin was employed in the present study to inhibit protein synthesis in dorsal root ganglion (DRG) neurons whose axons project into the lumbar nerve trunk of bullfrog tadpoles. The procedure was developed to examine, during tadpole metamorphosis, the ratio of fast-transported radiolabeled protein accumulating at the proximal side of a nerve trunk ligature to the total newly synthesized protein in the cell bodies of origin. The relatively small diameter and fragility of the developing lumbar nerve trunks necessitated introduction of ricin by bath application to the cut nerve end rather than by intraneural injection. Consistent uptake of ricin was achieved by pretreatment with the phospholipase A2 inhibitor, mepacrine, that blocks resealing of severed nerve fibers. Optimal time and dosage of ricin were established by determining the maximal achievable inhibition of [35S]methionine into DRG protein. In stage XVI tadpoles, maximal inhibition of approximately to 65% was observed after 16 h incubation in 2.5 mg/ml ricin. As evidence that neuronal protein synthesis was effectively suppressed, there was no detectable anterograde axonal transport of [35S]protein subsequent to ricin treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
Tang
H Z
HZ
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, CA 91010.
Tsai
A
A
Hammerschlag
R
R
D000818:Animals; D001345:Autoradiography; D001370:Axonal Transport / Q000502:physiology; D003864:Depression, Chemical; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology; D007150:Immunohistochemistry; D007814:Larva; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D011796:Quinacrine / Q000494:pharmacology; D011892:Rana catesbeiana; D012276:Ricin / Q000494:pharmacology*; D013462:Sulfur Radioisotopes
D009419:Nerve Tissue Proteins; D013462:Sulfur Radioisotopes; D012276:Ricin; D011796:Quinacrine