Do photobleached fluorescent microtubules move?: re-evaluation of fluorescence laser photobleaching both in vitro and in growing Xenopus axon.
We previously documented differences in the behavior of microtubules in growing axons of two types of neurons, adult mouse sensory neurons and Xenopus embryonal spinal cord neurons. Namely, the bulk of microtubules was stationary in mouse sensory neurons both by the method of photoactivation of caged-fluorescein-labeled tubulin and photobleaching of fluorescein-labeled tubulin, but the bulk of microtubules did translocate anterogradely by the method of photoactivation. Although these results indicated that the stationary nature of photobleached microtubules in mouse neurons is not an artifact derived from the high levels of energy required for the procedure, it has not yet been settled whether the photobleaching method can detect the movement of microtubules properly. Here we report photobleaching experiments on growing axons of Xenopus embryonal neurons. Anterograde movement of photobleached microtubules was observed at a frequency and translocation rate similar to the values determined by the method of photoactivation. Our results suggest that, under appropriate conditions, the photobleaching method is able to reveal the behavior of microtubules as accurately as the photoactivation method.
Okabe
S
S
Department of Anatomy and Cell Biology, School of Medicine, University of Tokyo, Japan.
Hirokawa
N
N
D000818:Animals; D001370:Axonal Transport; D001369:Axons / Q000648:ultrastructure*; D001692:Biological Transport; D002465:Cell Movement / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D007834:Lasers; D008870:Microtubules / Q000502:physiology / Q000528:radiation effects*; D010777:Photochemistry; D014404:Tubulin / Q000737:chemistry / Q000502:physiology*; D014982:Xenopus laevis
D005456:Fluorescent Dyes; D014404:Tubulin