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Journal of Neuroscience, Vol 7, 3245-3255, Copyright © 1987 by Society for Neuroscience
Reversible blockage of neurite development and growth cone formation in neuronal cultures of a temperature-sensitive mutant of Drosophila
YT Kim and CF Wu
Department of Biology, University of Iowa, Iowa City 52242.
It has been suggested that the membrane recycling process in the Drosophila
mutant shibirets1 (shits1) is reversibly blocked at restrictive temperature
(greater than 29 degrees C) in various cell types, including neurons, as a
result of enhanced vesicle fusion and disrupted membrane pinch-off (Poodry
and Edgar, 1979; Koenig et al., 1983; Kosaka and Ikeda, 1983a, b). We
analyzed the neuronal development of shits1 cells at the single-cell level
by using dissociated larval CNS culture. Compared to normal cultures, there
was a substantial reduction in neuronal adhesion to the substratum and a
retardation of growth cone formation in shits1 cultures at 30 degrees C.
Time-lapse studies showed that neurite initiation and elongation in shits1
cultures were suppressed at the restrictive temperature. Differentiated
shits1 neurons exhibited accelerated reabsorption and retarded arborization
of neurites at 30 degrees C. The above processes were reversible since
normal outgrowth was restored when the temperature was lowered. These
findings provide a new line of evidence that growth cone activity
determines neurite initiation, elongation, and branching. Furthermore, if
the primary defect of the shits1 mutation indeed resides in a membrane
recycling mechanism common to different cell types, our results would also
lend strong support to the notion that membrane recycling is crucial in the
regulation of neuronal membrane adhesion and growth cone activity.
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