Possible temperature‐dependent blockage of synaptic vesicle recycling induced by a single gene mutation in drosophila

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Abstract
The temperature-sensitive Drosophila mutant, shibirets1 (shi), has been shown to exhibit a reversible block in synaptic transmission at 30°C. Various synaptic terminals (neuromuscular, sensory, central) of this mutant were observed by electron microscopy. At 19°C, all terminals of shi showed essentially the same structural features as those of wild-type (Oregon-R) flies, while at 30°C (5 or 10 min of exposure) shi terminals exhibited various structural changes not seen in the wild type. The major structural change observed in all of the various types of terminals was the accumulation of many pitlike structures on the plasma membrane near presynaptic sites. These structures consisted of a spherical head portion, about 50–100 nm in diameter, and a cylindrical neck portion, about 20 nm long and 20–25 nm in diameter. The neck portion was surrounded by a kind of cytoplasmic dense material, about 10 nm thick, reminiscent of a “collar.” Thus, these pits are referred to as “collared pits.” Similar kinds of pits were observed, although very rarely, in wild-type flies at 19 and 30°C and in shi flies at 19°C. In addition, various degrees of vesicle depletion, and an increase in membranous structures (infoldings and cisternalike or tubulelike structures) often accompanied pit formation. The possibility that these pits are the result of a blocked step in the endocytotic process, which in turn causes vesicle depletion as exocytosis proceeds, is discussed.