Jung Jasmin, Loy Kristina, Schilling Eva-Maria, Röther Mareike, Brauner Jan M, Huth Tobias, Schlötzer-Schrehardt Ursula, Alzheimer Christian, Kornhuber Johannes, Welzel Oliver, Groemer Teja W
Department of Psychiatry and Psychotherapy, Friedrich-Alexander-University of Erlangen-Nuremberg, Schwabachanlage 6, 91054, Erlangen, Germany.
Mol Neurobiol. 2014 Apr;49(2):916-30. doi: 10.1007/s12035-013-8569-5. Epub 2013 Nov 5.
Effects of the antidepressant fluoxetine in therapeutic concentration on stimulation-dependent synaptic vesicle recycling were examined in cultured rat hippocampal neurons using fluorescence microscopy. Short-term administration of fluoxetine neither inhibited exocytosis nor endocytosis of RRP vesicular membranes. On the contrary, acute application of the drug markedly increased the size of the recycling pool of hippocampal synapses. This increase in recycling pool size was corroborated using the styryl dye FM 1-43, antibody staining with αSyt1-CypHer™5E and overexpression of synapto-pHluorin, and was accompanied by an increase in the frequency of miniature postsynaptic currents. Analysis of axonal transport and fluorescence recovery after photobleaching excluded vesicles originating from the synapse-spanning superpool as a source, indicating that these new release-competent vesicles derived from the resting pool. Super resolution microscopy and ultrastructural analysis by electron microscopy revealed that short-term incubation with fluoxetine had no influence on the number of active synapses and synaptic morphology compared to controls. These observations support the idea that therapeutic concentrations of fluoxetine enhance the recycling vesicle pool size and thus the recovery of neurotransmission from exhausting stimuli. The change in the recycling pool size is consistent with the plasticity hypothesis of the pathogenesis of major depressive disorder as stabilization of the vesicle recycling might be responsible for neural outgrowth and plasticity.
使用荧光显微镜在培养的大鼠海马神经元中研究了治疗浓度的抗抑郁药氟西汀对刺激依赖性突触小泡循环的影响。短期给予氟西汀既不抑制RRP囊泡膜的胞吐作用也不抑制其胞吞作用。相反,急性应用该药物显著增加了海马突触循环池的大小。使用苯乙烯基染料FM 1-43、αSyt1-CypHer™5E抗体染色和突触pHluorin的过表达证实了循环池大小的这种增加,并伴随着微小突触后电流频率的增加。轴突运输分析和光漂白后的荧光恢复排除了源自跨越突触的超级池的囊泡作为来源,表明这些新的具有释放能力的囊泡源自静息池。超分辨率显微镜和电子显微镜的超微结构分析表明,与对照组相比,短期用氟西汀孵育对活跃突触的数量和突触形态没有影响。这些观察结果支持这样一种观点,即氟西汀的治疗浓度会增加循环囊泡池的大小,从而增强神经传递从耗尽性刺激中恢复的能力。循环池大小的变化与重度抑郁症发病机制的可塑性假说是一致的,因为囊泡循环的稳定可能是神经生长和可塑性的原因。