Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release.
Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release.
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DOI：
10.1083/jcb.81.2.275
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发表时间：
1979-05
期刊：
影响因子：
--
通讯作者：
Evans L
中科院分区：
文献类型：
--
作者：
Heuser JE;Reese TS;Dennis MJ;Jan Y;Jan L;Evans L
我们描述了一种通过与冷金属块接触来冷冻生物组织的机器的设计和操作，该机器包含一个计时电路，该电路在冷冻前的最后几毫秒内刺激青蛙神经肌肉接头。我们展示了在递质放电过程中冷冻的神经末梢的冷冻断裂复制品，其显示胞吐作用中捕获的突触囊泡。我们使用4-氨基吡啶（4-AP），以增加每个神经冲动释放的递质量子的数量，并表明，被快速冷冻捕获的胞吐囊泡的数量急剧增加，表明一个囊泡经历胞吐为每个量子被释放。我们进行统计分析的空间分布的突触囊泡放电网站沿着的“活动区”，标志着这些神经的分泌区域，并显示，个别囊泡融合与质膜相互独立，预期从生理演示量子独立放电。因此，已经建立了快速冷冻作为捕获像突触传递一样短暂的生物过程的技术的效用。附录描述了一种新的电容方法来测量冻结速率，这表明我们的速冻技术的“时间分辨率”是2毫秒或更好。
We describe the design and operation of a machine that freezes biological tissues by contact with a cold metal block, which incorporates a timing circuit that stimulates frog neuromuscular junctions in the last few milliseconds before thay are frozen. We show freeze-fracture replicas of nerve terminals frozen during transmitter discharge, which display synpatic vesicles caught in the act of exocytosis. We use 4-aminopyridine (4-AP) to increase the number of transmitter quanta discharged with each nerve impulse, and show that the number of exocytotic vesicles caught by quick-freezing increases commensurately, indicating that one vesicle undergoes exocytosis for each quantum that is discharged. We perform statistical analyses on the spatial distribution of synaptic vesicle discharge sites along the "active zones" that mark the secretory regions of these nerves, and show that individual vesicles fuse with the plasma membrane independent of one another, as expected from physiological demonstrations that quanta are discharged independently. Thus, the utility of quick- freezing as a technique to capture biological processes as evanescent as synaptic transmission has been established. An appendix describes a new capacitance method to measure freezing rates, which shows that the "temporal resolution" of our quick-freezing technique is 2 ms or better.
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DOI：
10.1083/jcb.57.2.499
发表时间：
1973-05
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ceccarelli B;Hurlbut WP;Mauro A
通讯作者：
Mauro A
影响因子：
7.8
作者：
HEUSER, JE;REESE, TS
通讯作者：
REESE, TS
DOI：
10.1007/bf01098386
发表时间：
1974-01-01
期刊：
JOURNAL OF NEUROCYTOLOGY
影响因子：
--
作者：
BIRKS, RI
通讯作者：
BIRKS, RI
DOI：
10.1111/j.1365-2818.1976.tb02405.x
发表时间：
1976-01-01
期刊：
Journal of Microscopy (Oxford)
影响因子：
--
作者：
DEMPSEY G P;BULLIVANT S
通讯作者：
BULLIVANT S
影响因子：
5.5
作者：
BIRKS, R;HUXLEY, HE;KATZ, B
通讯作者：
KATZ, B