p75 neurotrophin receptor distribution and transport in cultured neurons.
In this work, we define a GFP-tagged version of the p75 neurotrophin receptor (p75GFP) as a useful molecular tool for studying its distribution and cellular dynamics. Expression and subcellular localization of p75GFP have been characterized in non-neuronal (HEK 293) and in neuronal (cortical and hippocampal) cells. By monitoring movements of intracellular p75GFP in living cultured hippocampal neurons, we found that the chimeric protein was transported by tubulo-vesicular structures both anterogradely (0.1-0.5microm/s) and retrogradely (0.1-1.1microm/s), with a faster component in retrogradely moving structures. Movements of the p75GFP-containing structures were inhibited by treatment with the microtubule-disrupting agent nocodazole. Our data indicate that p75GFP is a reliable tool for studying spatial and cellular properties of p75 in CNS neurons and that p75 transport inside neurons is mediated by microtubule-associated motors.
Formaggio
Elena
E
Department of Medicine and Public Health, Section Pharmacology, University of Verona, Policlinico G.B. Rossi, Verona, Italy. elena.formaggio@univr.it
Cantù
Cinzia
C
Chiamulera
Christian
C
Fumagalli
Guido F
GF
D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000378:metabolism / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D018715:Microscopy, Video; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D020409:Molecular Motor Proteins / Q000378:metabolism*; D009414:Nerve Growth Factors / Q000378:metabolism; D009419:Nerve Tissue Proteins; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D016716:PC12 Cells; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017978:Receptors, Growth Factor; D017475:Receptors, Nerve Growth Factor / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D022161:Transport Vesicles / Q000378:metabolism / Q000648:ultrastructure
D020409:Molecular Motor Proteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D017978:Receptors, Growth Factor; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; C068313:Ngfr protein, rat; D049452:Green Fluorescent Proteins