کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
6268327 1614628 2015 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Basic NeuroscienceVisualization of neurotransmitter uptake and release in serotonergic neurons
ترجمه فارسی عنوان
عصبشناسی پایه عصبی شدن جذب و انتشار نوروترانسمیتر در نورونهای سروتونرژیک
موضوعات مرتبط
علوم زیستی و بیوفناوری علم عصب شناسی علوم اعصاب (عمومی)
چکیده انگلیسی


- We have established the visualization of fluorescent transmitter-like substrates in stem cell-derived serotonergic neurons.
- We have combined uptake and release of fluorescent substrates into and from serotonergic neurons with quantitative image analysis.
- These methods allow optical discrimination of axonal from somatodendritic transmitter release.

BackgroundTo study serotonergic volume neurotransmission at cellular level it needs to investigate neurotransmitter release and re-uptake sites in serotonergic neurons. However, due to the low number of cell bodies in the raphe nuclei and their widely branching neurites, serotonergic neuronal cultures are not accessible ex vivo.New methodWe have combined differentiation protocols for the generation of stem cell-derived serotonergic neurons together with confocal microscopy to study the uptake and release of fluorescent substrates known to be selectively taken up by monoaminergic neurons. These substances include: (i) 4-(4-(dimethylamino)styryl)-N-methylpyridiunium (ASP+), an analog of the neurotoxin MPP+; (ii) the fluorescent false neurotransmitter (FFN511); and (iii) serotonin (5-hydroxytryptamine; 5-HT) itself, which is known to emit fluorescence upon excitation at 320-460 nm.ResultASP+ is taken up into living serotonergic neurons through the serotonin transporter, but not accumulated into synaptic vesicles; FFN511 diffuses in a SERT-independent way into serotonergic neurons and accumulated into synaptic vesicles. KCl-induced release of FFN511 and 5-HT can be visualized and quantified in living serotonergic neurons.Comparison with existing methodsApplication of ASP+ so far has been used to investigate substrate/transporter interactions; studies on FFN511 uptake and release have only been performed in dopaminergic neurons; quantitative studies on uptake and release of 5-HT in living serotonergic neurons have not been reported yet.ConclusionThe differentiation protocols for the generation of stem cell-derived serotonergic neurons combined with the application of different fluorescent dyes allow to quantify neurotransmitter uptake and release in living serotonergic neurons in vitro.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Neuroscience Methods - Volume 241, 15 February 2015, Pages 10-17
نویسندگان
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