title: Time course and temperature dependence of the membrane translocation of tetanus and botulinum neurotoxins C and D in neurons.
abstract: Tetanus and botulinum neurotoxins act inside nerve terminals and, therefore, they have to translocate across a membrane to reach their targets. This translocation is driven by a pH gradient, acidic on the cis side and neutral on the cytosol. Recently, a protocol to induce translocation from the plasma membrane was established. Here, we have used this approach to study the temperature dependence and time course of the entry of the L chain of tetanus neurotoxin and of botulinum neurotoxins type C and D across the plasma membrane of cerebellar granular neurons. The time course of translocation of the L chain varies for the three neurotoxins, but it remains in the range of minutes at 37 °C, whilst it takes much longer at 20 °C. BoNT/C does not enter neurons at 20 °C. Translocation also depends on the dimension of the pH gradient. These data are discussed with respect to the contribution of the membrane translocation step to the total time to paralysis and to the low toxicity of these neurotoxins in cold-blood vertebrates.
authors.affiliation: Dipartimento di Scienze Biomediche and Istituto CNR di Neuroscienze, Università di Padova, Viale G. Colombo 3, 35131 Padova, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Pirazzini
authors.forename: Ornella
authors.initials: O
authors.lastname: Rossetto
authors.forename: Cristina
authors.initials: C
authors.lastname: Bertasio
authors.forename: Fulvio
authors.initials: F
authors.lastname: Bordin
authors.forename: Clifford C
authors.initials: CC
authors.lastname: Shone
authors.forename: Thomas
authors.initials: T
authors.lastname: Binz
authors.forename: Cesare
authors.initials: C
authors.lastname: Montecucco
mesh_terms: D000818:Animals; D001905:Botulinum Toxins / Q000378:metabolism* / Q000633:toxicity; D002462:Cell Membrane / Q000201:enzymology*; D002478:Cells, Cultured; D006863:Hydrogen-Ion Concentration; D008666:Metalloendopeptidases / Q000378:metabolism* / Q000633:toxicity; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D014176:Protein Biosynthesis; D051381:Rats; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism; D013696:Temperature; D013744:Tetanus Toxin / Q000378:metabolism* / Q000633:toxicity; D013997:Time Factors
chemical_list: D050825:Synaptosomal-Associated Protein 25; D013744:Tetanus Toxin; C035979:tetanospasmin; C096325:botulinum toxin type D; D008666:Metalloendopeptidases; D001905:Botulinum Toxins; C096324:botulinum toxin type C