Viancour T A, Sheller R A, Bittner G D, Seshan K R
University of Maryland, Department of Biological Sciences, Catonsville 21228.
Brain Res. 1988 Jan 26;439(1-2):211-21. doi: 10.1016/0006-8993(88)91477-1.
Segmental lateral giant axons (SLGAs) in crayfish were used to determine whether functionally intact proteins can move between axons under physiological conditions. Horseradish peroxidase (HRP) was chosen as the tracer protein because its localization requires intact enzymatic activity and because it can be localized in living cells using a non-cytotoxic procedure. Following iontophoretic injection of HRP in a single SLGA, HRP often transferred to adjacent SLGAs. HRP transferred from an injected SLGA to a caudal SLGA with greater frequency than HRP transferred to a rostral SLGA. When HRP transferred between SLGAs, it was ultrastructurally associated with vesicles on both sides of septate junctions between adjacent SLGAs and was also seen in the perijunctional extracellular space. There was no difference between the electrical resistance of synapses at which HRP transferred and those synapses where HRP did not transfer. HRP transfer was significantly reduced when axons were bathed in reduced calcium saline. These and other observations indicate that axon-to-axon transport in this system is accomplished by exocytosis of HRP from injected axons followed by its endocytotic uptake by adjacent, non-injected axons. Similar transfer of endogenous proteins may contribute to the long-term survival for months to years of distal stumps of severed SLGAs.
小龙虾的节段性外侧巨轴突(SLGA)被用于确定功能完整的蛋白质在生理条件下是否能在轴突之间移动。选择辣根过氧化物酶(HRP)作为示踪蛋白,是因为其定位需要完整的酶活性,并且可以使用非细胞毒性程序在活细胞中定位。在单个SLGA中通过离子电渗法注射HRP后,HRP经常转移到相邻的SLGA。HRP从注射的SLGA转移到尾侧SLGA的频率高于转移到头侧SLGA的频率。当HRP在SLGA之间转移时,在超微结构上它与相邻SLGA之间的分隔连接两侧的囊泡相关联,并且在连接周细胞外空间中也可见。HRP转移的突触与未转移HRP的突触之间的电阻没有差异。当轴突浸泡在低钙盐溶液中时,HRP转移显著减少。这些以及其他观察结果表明,该系统中的轴突到轴突运输是通过注射轴突中HRP的胞吐作用,随后被相邻的未注射轴突通过胞吞作用摄取来完成的。内源性蛋白质的类似转移可能有助于切断的SLGA远端残端存活数月至数年。